scholarly journals Examination of the minimum inhibitory concentration of amoxicillin and marbofloxacin against Streptococcus suis using standardised methods

2020 ◽  
Vol 65 (No. 9) ◽  
pp. 377-386
Author(s):  
D Sperling ◽  
H Karembe ◽  
M Zouharova ◽  
K Nedbalcova
Keyword(s):  
Inhibitory Concentration ◽  
Streptococcus Suis ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Microdilution Method ◽  
Linear Relationships ◽  
Broth Microdilution Method ◽  
Significant Difference ◽  
Agar Dilution ◽  
Good Agreement

The results of the antimicrobial susceptibility testing of clinical isolates Streptococcus suis to amoxicillin and marbofloxacin obtained by the agar dilution method and broth microdilution method with the results obtained by the commercially available E-test were compared. Comparisons between the methods based on the determination of the minimal inhibitory concentration (MIC) of the antimicrobials were assessed based on the degree and frequency of the categorical agreement (Agar dilution method as a reference system) and the percentage of the categorical agreement and error rate. A statistical evaluation was determined using the Bland-Atman method. The presented MIC values, determined for the isolates in the E-test, were slightly different from the MIC values determined by the dilution tests, mainly due to the different defined testing concentrations. For the E-test as the test system and agar-dilution method as the reference system, no error of any class was detected (very major, major and minor error) and a complete categorical agreement was obtained between the evaluated methods for amoxicillin. For amoxicillin, the regression and correlation analysis show linear relationships between the E-test and the two dilution methods with significant coefficients of determination (0.62 and 0.75). The slopes of the equality and regression lines were not significantly different. However, the E-test tends to slightly overestimate the MIC values when compared to the microdilution. The reverse is true when compared with the agar dilution. There was good agreement between the E-test and the dilution methods with a low bias (0.001 3 and −0.005 0), all the experimental data were within the computed limits of agreement. For marbofloxacin, the same trends were observed with lower coefficients of determination (0.42 and 0.73) and a less favourable agreement. The E-test constantly underestimated the MIC values when compared to the two dilution methods. No significant difference between the microdilution and agar dilution was obtained.

scholarly journals Comparison of Broth Microdilution Method UsingHaemophilus Test Medium and Agar Dilution Method for Susceptibility Testing of Eikenella corrodens

1998 ◽  
Vol 36 (8) ◽  
pp. 2386-2388 ◽  
Author(s):  
Luis Alcalá ◽  
Fernando García-Garrote ◽  
Emilia Cercenado ◽  
Teresa Peláez ◽  
Gema Ramos ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Broth Microdilution ◽  
Test Medium ◽  
Eikenella Corrodens ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Agar Dilution

Susceptibility testing of Eikenella corrodens is usually performed by a Mueller-Hinton sheep blood agar dilution (AD) method. However, this method is impractical for testing only a few strains. We compared AD with the broth microdilution method usingHaemophilus test medium (HTM) in order to determine the susceptibility of 36 clinical isolates of E. corrodens to eight antimicrobial agents. MICs obtained by the HTM method yielded 95.5 and 84% agreement (within 2 and 1 log2 dilutions, respectively) with those obtained by AD. The HTM method with incubation in CO2 for 48 h was highly reproducible and constitutes an easy alternative for antimicrobial susceptibility testing of E. corrodens.

scholarly journals Validation of a method of broth microdilution for the determination of antibacterial activity of essential oils

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Vanegas David ◽  
Abril-Novillo Andrea ◽  
Khachatryan Aleksandr ◽  
Jerves-Andrade Lourdes ◽  
Peñaherrera Eugenia ◽  
...  
Keyword(s):  
Essential Oils ◽  
Sensitivity And Specificity ◽  
Inhibitory Concentration ◽  
Tween 80 ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Broth Microdilution ◽  
Continuous Contact ◽  
Microdilution Method ◽  
Broth Microdilution Method

Abstract Objective The aim of the present study was to adapt and optimize a broth microdilution method and compare it to the agar dilution method for the evaluation of activity of essential oils from medicinal plants against Gram-negative bacteria. Based on bibliographic research, active and not active oils were selected. The sensitivity and specificity were established as parameters for validation. The comparison between both methods was made using contingency analysis tables, based on the observed frequencies. For both methods, the minimum inhibitory concentration was determined against Escherichiacoli strains, in an essential oil concentration range between 0.03 and 0.48% (v/v). Results A stable emulsion formation was achieved with the addition of Tween 80 and constant agitation, guaranteeing the continuous contact of oil with bacteria (critical step in the microdilution method). The statistical analysis of results obtained with both methods presented a good sensitivity and specificity (100% in both cases), which let us correctly discriminate between active and non-active oils. The values obtained for the minimal inhibitory concentration were independent of the technique used. Finally, the obtained results show that the validated microtechnique allows important diminishment of time and resources for investigations dealing with essential oils or lipophilic extracts 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 Gemifloxacin versus Five Quinolones and Two Macrolides against 271 Spanish Isolates of Legionella pneumophila: Influence of Charcoal on Susceptibility Test Results

2000 ◽  
Vol 44 (8) ◽  
pp. 2176-2178 ◽  
Author(s):  
M. T. García ◽  
C. Pelaz ◽  
M. J. Giménez ◽  
L. Aguilar
Keyword(s):  
Legionella Pneumophila ◽  
Free Drug ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Test Results ◽  
Progressive Decrease ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
Agar Dilution

ABSTRACT The MICs at which 90% of isolates are inhibited for gemifloxacin, trovafloxacin, and grepafloxacin were low (≤0.01 μg/ml) for 271Legionella isolates when they were determined by the broth microdilution method but increased (≥6 dilutions) when they were determined by the agar dilution method. This was due to the charcoal in the agar dilution medium, as shown by the progressive decrease in the MICs when the charcoal concentrations decreased. As free drug is the active fraction, charcoal binding should be considered.

scholarly journals Drug resistance of yeasts isolated from oropharyngeal candidiasis in aids patients

1998 ◽  
Vol 29 (4) ◽  
pp. 271-275
Author(s):  
Maria do Rosário Rodrigues Silva ◽  
Claudete Rodrigues de Paula ◽  
Soraya Cristina Silva ◽  
Théo Rodrigues Costa ◽  
Márcio Rodrigues Costa
Keyword(s):  
Amphotericin B ◽  
High Frequency ◽  
Inhibitory Concentration ◽  
Serum Levels ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Oropharyngeal Candidiasis ◽  
Candida Spp ◽  
Aids Patients ◽  
Agar Dilution

Candida spp was isolated from 59 (68.60%) out of eighty six samples of oral mucosa of AIDS patients. The identification, based or the production of a germ tube and chlamydospores, and on the assimilation and fermentation of carbohydrates, revealed 52 strains (88.13%) of C. albicans, 4 (6.77%) of C. tropicalis and 3 (5.08%) of C. krusei. The susceptibility of these strains to amphotericin B, flucytosine, itraconazole, fluconazole and ketoconazole was determined using the agar dilution method. Comparing the minimum inhibitory concentration values found in the susceptibility test with the serum levels achieved by these drugs, only 8.47% and 5.08% of the yeasts strains proved to be resistant to amphotericin B and flucitosyne, respectively. A high frequency of strains resistant to azole derivatives (25.42%, to itraconazole, 45.76%, to ketoconazole and 66.10% to fluconazole) was observed.

scholarly journals Comparison of methods for the determination of antimicrobial resistance in Campylobacter spp. human and the food chain isolates

2008 ◽  
Vol 52 (No. 4) ◽  
pp. 169-174
Author(s):  
M. Holasova ◽  
R. Karpiskova ◽  
S. Karpiskova ◽  
V. Babak ◽  
J. Schlegelova
Keyword(s):  
Nalidixic Acid ◽  
Antimicrobial Agents ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Disk Diffusion Method ◽  
Microdilution Method ◽  
Study Results ◽  
Agar Dilution

With a microdilution method, using the commercial diagnostic test Sensititre Susceptibility Plates for Campylobacter MIC (Trek Diagnostic Systems, Cleveland, OH, USA), disk diffusion and agar dilution method, resistance to six antimicrobial agents were examined in a reference strain <i>Campylobacter jejuni</i> ATCC 33560 and 73 thermo-tolerant isolates of <i>Campylobacter</i> spp. For the microdilution method and all tested antimicrobial agents, our determined values of microbiological breakpoints of resistant strains were suggested as the minimum inhibitory concentration (MIC<sub>R</sub>) for ciprofloxacin &ge; 0.5, erythromycin &ge; 4, gentamicin &ge; 4, nalidixic acid &ge; 32 and tetracycline &ge; 4 &mu;g/ml. On the basis of our study results, strains resistant to clindamycin were MIC<sub>R</sub> &ge; 2 &mu;g/ml for the dilution methods and a zone diameter R ≤ 16 mm for the disk diffusion method. Comparison of the results of the resistance examination, a microdilution method and disk diffusion method with the reference agar dilution method, showed that all compared methods yielded identical results with the exception of the resistance determination in erythromycin and nalidixic acid. The errors were mostly the result of the interpretation criteria for MIC<sub>R</sub> of agar dilution method and different conditions of cultivation used. However, the compared methods, provide results comparable with the reference method having greater convenience of measurement.

scholarly journals Characterization of Clinical Isolates ofEnterococciwith Special Reference to Glycopeptide Susceptibility at a Tertiary Care Center of Eastern Nepal

2019 ◽  
Vol 2019 ◽  
pp. 1-8 ◽  
Author(s):  
Aasish Karna ◽  
Ratna Baral ◽  
Basudha Khanal
Keyword(s):  
Multidrug Resistance ◽  
Minimum Inhibitory Concentration ◽  
Antimicrobial Susceptibility ◽  
Inhibitory Concentration ◽  
Tertiary Care ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Disc Diffusion ◽  
Diffusion Technique ◽  
Agar Dilution

Background. Enterococci, once considered as a harmless commensal of intestine, have now emerged as medically important pathogens and are associated with both community-acquired and nosocomial infections. They bear the potential to exhibit resistance against all commonly used antibiotics either by inherent or acquired mechanism, posing a therapeutic challenge.Objectives. This study aimed to characterize enterococci up to the species level and study their antibiogram with special regard to vancomycin.Methods. A descriptive cross-sectional study was conducted in the Department of Microbiology, B.P. Koirala Institute of Health Sciences, Dharan, Nepal, from February to May 2017. A total of 91 enterococcal isolates recovered from clinical specimens were investigated in this study. Their identification and speciation were done according to standard microbiological guidelines. Kirby–Bauer disc diffusion technique was used to study antimicrobial susceptibility pattern, whereas minimum inhibitory concentration of vancomycin was determined by the agar dilution method, with reference to Clinical and Laboratory Standards Institute guidelines.Results. Seven different species of enterococci were isolated,E. faecalisandE. faeciumaccounting about 45% each. The other species encountered wereE.avium,E.cecorum,E.dispar,E.durans, andE.raffinosus. Highest proportion of antimicrobial susceptibility was recorded for linezolid (97.8%), followed by teicoplanin (95.6%) and high-level gentamicin (81.3%). Sensitivity to vancomycin was seen in 79.1% isolates. Likewise, 82.1% of urinary strains were susceptible to nitrofurantoin. A total of 4 disparities were observed between the disc diffusion technique and agar dilution method in determining vancomycin resistance. Multidrug resistance was observed in 31.9% isolates. The overall prevalence of vancomycin-resistant enterococci based on the standard minimum inhibitory concentration method was 25.3%.Conclusions.Enterococcus faecalisandE. faeciumwere the predominant species in causing enterococcal infections. The alarming rise in prevalence of vancomycin and multidrug resistance strains warrants immediate, adequate, and efficient surveillance program to prevent and control its spread.

Antibiotic Resistance of Salmonella Isolated from Hog, Beef, and Chicken Carcass Samples from Provincially Inspected Abattoirs in Ontario

2004 ◽  
Vol 67 (3) ◽  
pp. 448-455 ◽  
Author(s):  
C. LARKIN ◽  
C. POPPE ◽  
B. MCNAB ◽  
B. MCEWEN ◽  
A. MAHDI ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
The United States ◽  
Test Methods ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Chicken Carcass ◽  
Resistance Patterns ◽  
Agar Dilution ◽  
Microbroth Dilution ◽  
Good Agreement

The emergence of antimicrobial-resistant Salmonella organisms, especially Salmonella Typhimurium DT104, has been reported in many countries, including the United States and Canada. The purposes of this study were to determine the antimicrobial resistance patterns of Salmonella isolated from hog, beef, and chicken carcasses from provincially inspected abattoirs in Ontario and to determine the agreement between the agar dilution method and the microbroth dilution method for measurement of antimicrobial resistance of the isolates. Antimicrobial resistance of Salmonella isolates from hogs (n = 71), beef (n = 24), and chicken (n = 295) to amikacin, ampicillin, cephalothin, chloramphenicol, ciprofloxacin, gentamicin, streptomycin, sulfamethoxazole,and tetracycline was determined using the two methods. None of the 390 isolates were resistant to ciprofloxacin at levels of 0.125 μg/ml. All chicken and hog isolates were sensitive to amikacin, whereas all beef isolates were sensitive to both amikacin and gentamicin. Multiple antimicrobial resistance (resistance to more than one antimicrobial) was found in 29% of bovine isolates and 42% of porcine isolates using both methods for testing and in 42% by the agar dilution and 33% by the microbroth dilution methods in the chicken isolates. Overall, there was good agreement between the two test methods for resistance to most of the antimicrobials, with disagreement found in the results in 1.3% of the isolates for ampicillin and sulfamethoxazole, 8.2% for streptomycin, 5.6% for cephalothin, and 1.0% of the isolates for tetracycline. The lack of agreement between the two test methods was found mostly among the chicken isolates.

scholarly journals In VitroActivity of Fosfomycin against a Collection of Clinical Pseudomonas aeruginosa Isolates from 16 Spanish Hospitals: Establishing the Validity of Standard Broth Microdilution as Susceptibility Testing Method

2013 ◽  
Vol 57 (11) ◽  
pp. 5701-5703 ◽  
Author(s):  
María Díez-Aguilar ◽  
María-Isabel Morosini ◽  
Rosa del Campo ◽  
María García-Castillo ◽  
Javier Zamora ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Susceptibility Testing ◽  
Disk Diffusion ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
Broth Microdilution ◽  
Content Type ◽  
Testing Method ◽  
Broth Microdilution Method ◽  
Agar Dilution

ABSTRACTThe broth microdilution method for fosfomycin andPseudomonas aeruginosawas assessed and compared with the approved agar dilution method in 206 genetically unrelatedP. aeruginosaclinical isolates. Essential agreement between the two methods was 84%, and categorical agreement was 89.3%. Additionally, Etest and disk diffusion assays were performed. Results validate broth microdilution as a reliable susceptibility testing method for fosfomycin againstP. aeruginosa. Conversely, unacceptable concordance was established between Etest and disk diffusion results with agar dilution results.

scholarly journals Antimicrobial resistance of Enterococcus faecium and Enterococcus faecalis, isolated from blood culture of patients with hematological malignancies during different study periods

2021 ◽  
Vol 16 (1) ◽  
pp. 54-63
Author(s):  
A. V. Fedorova ◽  
G. A. Klyasova ◽  
I. N. Frolova ◽  
S. A. Khrulnova ◽  
A. V. Vetokhina ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Blood Culture ◽  
Enterococcus Faecalis ◽  
Enterococcus Faecium ◽  
Dilution Method ◽  
Agar Dilution Method ◽  
High Susceptibility ◽  
Microdilution Method ◽  
Broth Microdilution Method ◽  
High Level

Objective: to determine antimicrobial resistance of Enterococcus faecium and Enterococcus faecalis isolated from blood culture of hematological patients during different study periods.Materials and methods. Antimicrobial susceptibility of Enterococcus spp., collected as part of the multicenter study was tested by the broth microdilution method (USA Clinical and Laboratory Standards Institute (CLSI), 2018), to daptomycin by Etest (bioMeriéux, France). High-level gentamicin resistance (HLGR) and high-level streptomycin resistance (HLSR) was performed by the agar dilution method (CLSI (Oxoid, UK), 2018).Results. The susceptibility of 366 E. faecium (157 in 2002-2009 and 209 in 2010-2017) and 86 E. faecalis (44 in 20022009 and 42 in 2010-2017) was studied. In the second study period (2010-2017) the rise of vancomycin-resistant E. faecium (VREF) increased from 8.3 % to 23.4 % (p = 0.0001), and two linezolid-resistant (LREF) were identified. All VREF and LREF remained susceptible to daptomycin and tigecycline. The rate of susceptible to tetracycline E. faecium remained the same (73.9 and 74.6 %), and an increase in susceptibility to chloramphenicol (74.5 and 82.3 %) was observed. Susceptibility of E. faecium to tetracycline was detected with almost the same rate and in a part of isolates, the increase of susceptibility to chloramphenicol was registered during the analyzed periods. The rise of E. faecium susceptible to HLGR and HLSR has increased significantly in 2010-2017 compared to 2002-2009. Erythromycin, levofloxacin, ampicillin and penicillin had the least activity against E. faecium (less than 5 %).All E. faecalis were susceptible to tigecycline, linezolid, and teicoplanin. Only one of E. faecalis had intermediate resistance to vancomycin. High susceptibility to ampicillin in E. faecalis remained unchanged (97.7 and 97.6 %, respectively). In the second period of the study the rise of susceptible E. faecalis decreased significantly to penicillin (from 97.7 % to 76.2 %), to levofloxacin (from 59.1 % to 31 %), to HLSR (from 52.3 % до 31 %), and to HLGR (from 47.7 % to 26.2 %), remained unchanged to chloramphenicol (52.3 % and 50 %) and was minimal to erythromycin and tetracycline.Conclusion. The study demonstrated higher rates of antibiotic resistance among E. faecium, which consisted of an increase in VREF and the appearance of linezolid-resistant strains. High susceptibility to ampicillin remained in E. faecalis, but there was an increase in resistance to penicillin and aminoglycosides.

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