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 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.

In vitro activity of a novel antibacterial agent, levonadifloxacin, against clinical isolates collected in a prospective, multicentre surveillance study in India during 2016–18

2019 ◽  
Vol 75 (3) ◽  
pp. 600-608 ◽  
Author(s):  
Boppe Appalaraju ◽  
Sujata Baveja ◽  
Shrikala Baliga ◽  
Suchitra Shenoy ◽  
Renu Bhardwaj ◽  
...  
Keyword(s):  
Therapeutic Option ◽  
Tertiary Care ◽  
Vitro Activity ◽  
Clinical Isolates ◽  
Surveillance Study ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
In Vitro Activity ◽  
Broth Microdilution Method

Abstract Background Levonadifloxacin is a novel antibiotic belonging to the benzoquinolizine subclass of fluoroquinolones with potent activity against MRSA and quinolone-resistant Staphylococcus aureus. IV levonadifloxacin and its oral prodrug alalevonadifloxacin have recently been approved in India for the treatment of acute bacterial skin and skin structure infections (ABSSSIs) including diabetic foot infections. Objectives To investigate the in vitro activity of levonadifloxacin against contemporary clinical isolates collected from multiple tertiary care hospitals across India in the Antimicrobial Susceptibility Profiling of Indian Resistotypes (ASPIRE) surveillance study. Methods A total of 1376 clinical isolates, consisting of staphylococci (n = 677), streptococci (n = 178), Enterobacterales (n = 320), Pseudomonas aeruginosa (n = 140) and Acinetobacter baumannii (n = 61), collected (2016–18) from 16 tertiary hospitals located across 12 states in India, were included in the study. The MICs of levonadifloxacin and comparator antibiotics were determined using the reference agar dilution method and broth microdilution method. Results Levonadifloxacin exhibited potent activity against MSSA (MIC50/90: 0.5/1 mg/L), MRSA (MIC50/90: 0.5/1 mg/L) and levofloxacin-resistant S. aureus (MIC50/90: 1/1 mg/L) isolates. Similarly, potent activity of levonadifloxacin was also observed against CoNS including MDR isolates (MIC50/90: 1/2 mg/L). Against Streptococcus pneumoniae, levonadifloxacin (MIC50/90: 0.5/0.5 mg/L) showed superior activity compared with levofloxacin (MIC50/90: 1/2 mg/L). Among levofloxacin-susceptible Enterobacterales, 80.6% of isolates were inhibited at ≤2 mg/L levonadifloxacin. Conclusions Levonadifloxacin displayed potent activity against contemporary MRSA and fluoroquinolone-resistant staphylococcal isolates, thus offering a valuable IV as well as an oral therapeutic option for the treatment of ABSSSIs. Furthermore, levonadifloxacin exhibited a broad-spectrum activity profile as evident from its activity against streptococci and levofloxacin-susceptible Gram-negative isolates.

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 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.

scholarly journals Problems Related to Determination of MICs of Oximino-Type Expanded-Spectrum Cephems for Proteus vulgaris

2000 ◽  
Vol 38 (2) ◽  
pp. 677-681
Author(s):  
Akira Ohno ◽  
Yoshikazu Ishii ◽  
Ling Ma ◽  
Keizo Yamaguchi
Keyword(s):  
Dilution Method ◽  
Agar Dilution Method ◽  
Proteus Vulgaris ◽  
In Vitro Susceptibility ◽  
Microdilution Method ◽  
Defective Mutant ◽  
Broth Microdilution Method ◽  
Agar Dilution ◽  
Isogenic Mutants

During in vitro susceptibility testing of clinical isolates ofProteus vulgaris, we noted that the MICs of several expanded-spectrum cephems were much higher in the broth microdilution method than in the agar dilution method (termed the MIC gap phenomenon). Here we investigated the mechanism of the MIC gap phenomenon. Cephems with the MIC gap phenomenon were of the oximino type, such as cefotaxime, cefteram, and cefpodoxime, which serve as good substrates for inducible class A β-lactamase (CumA) enzymes produced by P. vulgaris; this finding suggests a relationship between the MIC gap phenomenon and CumA. Since peptidoglycan recycling shares a system common to that inducing CumA, we analyzed the mechanism of the MIC gap phenomenon using P. vulgaris B317 and isogenic mutants with mutations in the peptidoglycan recycling and β-lactamase induction systems. The MIC gap phenomenon was observed in the parent strain B317 but not in B317G (cumG-defective mutant; defective peptidoglycan recycling) and B317R (cumR-defective mutant; defective CumA transcriptional regulator). No β-lactamase activity was detected in B317G and B317R. β-Lactamase activity and the MIC gap phenomenon were restored in B317G/pMD301 (strain transcomplemented by a clonedcumG gene) and B317R/pMD501 (strain transcomplemented by a cloned cumR gene). MICs determined by the agar dilution method increased when lower agar concentrations were used. Our results indicated that the mechanism of the MIC gap phenomenon is related to peptidoglycan recycling and CumA induction systems. However, it remains unclear how β-lactamase induction of P. vulgaris is suppressed on agar plates.

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 Antibiotic Susceptibility Pattern ofMycobacterium marinum

2000 ◽  
Vol 44 (11) ◽  
pp. 3133-3136 ◽  
Author(s):  
Alexandra Aubry ◽  
Vincent Jarlier ◽  
Sylvie Escolano ◽  
Chantal Truffot-Pernot ◽  
Emmanuelle Cambau
Keyword(s):  
Antibiotic Susceptibility ◽  
Geometric Mean ◽  
Multidrug Resistant ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Susceptibility Pattern ◽  
Cutaneous Infections ◽  
Agar Dilution

ABSTRACT In vitro activities of 17 antibiotics against 53 clinical strains of Mycobacterium marinum, an atypical mycobacterium responsible for cutaneous infections, were determined using the reference agar dilution method. Rifampin and rifabutin were the most active drugs (MICs at which 90% of the isolates tested were inhibited [MIC90s], 0.5 and 0.6 μg/ml, respectively). MICs of minocycline (MIC90, 4 μg/ml), doxycycline (MIC90, 16 μg/ml), clarithromycin (MIC90, 4 μg/ml), sparfloxacin (MIC90, 2 μg/ml), moxifloxacin (MIC90, 1 μg/ml), imipenem (MIC90, 8 μg/ml), sulfamethoxazole (MIC90, 8 μg/ml) and amikacin (MIC90, 4 μg/ml) were close to the susceptibility breakpoints. MICs of isoniazid, ethambutol, trimethoprim, azithromycin, ciprofloxacin, ofloxacin, and levofloxacin were above the concentrations usually obtained in vivo. For each drug, the MIC50, geometric mean MIC, and modal MIC were very close, showing that all the strains had a similar susceptibility pattern. Percent agreement (within ±1 log2 dilution) between MICs yielded by the Etest method and by the agar dilution method used as reference were 83, 59, 43, and 24% for minocycline, rifampin, clarithromycin, and sparfloxacin, respectively. Reproducibility with the Etest was low, in contrast to that with the agar dilution method. In conclusion, M. marinum is a naturally multidrug-resistant species for which the agar dilution method is more accurate than the Etest for antibiotic susceptibility testing.

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.

scholarly journals In Vitro and In Vivo Evaluations of β-Lactam/β-Lactamase Mono- and Combined Therapies against Carbapenem-Nonsusceptible Enterobacteriaceae in Taiwan

2020 ◽  
Vol 8 (12) ◽  
pp. 1981
Author(s):  
Tsung-Ying Yang ◽  
Ya-Ju Hsieh ◽  
Li-Ting Kao ◽  
Guan-Hong Liu ◽  
Shao-Hsuan Lian ◽  
...  
Keyword(s):  
Carbapenem Resistance ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Geographic Variations ◽  
C Elegans ◽  
Agar Dilution ◽  
Antibiotic Susceptibility Patterns ◽  
Resistance Rates

Increasing carbapenem resistance rates worldwide underscored the urgent need of novel antimicrobials. Ceftazidime–avibactam and aztreonam–avibactam combinations are developed to combat carbapenem resistance, but biological and geographic variations must be considered for antibiotic susceptibility patterns varied. Thus, we sought to assess the susceptibilities of ceftazidime–avibactam and aztreonam–avibactam against 660 carbapenem-nonsusceptible Enterobacteriaceae isolates (472 Klebsiella pneumoniae and 188 Escherichia coli) collected during an earlier Taiwan surveillance study. Agar dilution method was used to determine ceftazidime–avibactam and aztreonam–avibactam susceptibility. Metallo-carbapenemase’s contribution to resistance were investigated with EDTA addition. The in vivo efficacies were evaluated using a Caenorhabditis elegans model. High susceptibility rates were observed for ceftazidime–avibactam and aztreonam–avibactam against the 472 carbapenem-nonsusceptible K. pneumoniae (CnsKP) (85.2% and 95.3%, respectively) and 188 carbapenem-nonsusceptible E. coli (CnsEC) isolates (91.5% and 94.1%, respectively). For non-metallo-carbapenemase producers, the susceptibility rates for ceftazidime–avibactam were 93.6% for CnsKP and 97.7% for CnsEC, whereas only 7.1% CnsKP and 11.1% CnsEC in metallo-carbapenemase producers were susceptible to ceftazidime–avibactam. Of all isolates, 95.3% CnsKP and 94.1% CnsEC were susceptible to aztreonam–avibactam. In C. elegans model, ceftazidime–avibactam and aztreonam–avibactam revealed effective against a blaKPC-producing K. pneumoniae isolate in vivo. Our results propose a positive therapeutic approach for both combinations against carbapenem-nonsusceptible Enterobacteriaceae in Taiwan.

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