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      In vitro synergistic activity of antibiotic combinations against Brucella melitensis using E-test methodology

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          Abstract

          The treatment of brucellosis is still problematic, because of high rates of treatment failure or relapses. As the microorganism is an intracellular pathogen, treatment requires combined regimens. However, limited existing data on in vitro combinations are avaliable for Brucellae. The aim of this study was to investigate the in vitro efficacy of various traditional and new antibiotic combinations against 16 Brucella melitensis strains. The combination effect of antimicrobial agents was evaluated by E-test synergy method to obtain a fractional inhibitory concentration (FIC) index. Co-Trimoxazole (SXT) and moxifloxocin (MXF) exhibited the lowest MIC, while Rifampin (RIF) had the highest MIC in the study. Combinations with RIF showed the best synergistic activity (100% of RIF-tetracycline (TET), and 87.5% of RIF-SXT). Synergistic activity was also detected at seven (43.7%) of ciprofloxocin (CIP)-SXT, four (25%) of TET-MXF, and two (12.5%) of TET-SXT combinations. The combinations that demonstrated additivity were TET-SXT, CIP-SXT and TET-MXF. Antagonism was observed only with the TET-Streptomycin (STR) combination in three strains (18.8%). Further work including randomized controlled clinical trials is required to fully evaluate the usefulness of these data.

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          Comparison of three different in vitro methods of detecting synergy: time-kill, checkerboard, and E test.

          R. L. . White, D. Burgess, M Manduru … (1996)
          An in vitro method of detecting synergy which is simple to perform, accurate, and reproducible and has the potential for clinical extrapolation is desirable. Time-kill and checkerboard methods are the most widely used techniques to assess synergy but are time-consuming and labor-intensive. The Epsilometer test (E test), a less technically demanding test, has not been well studied for synergy testing. We performed synergy testing of Escherichia coli ATCC 35218, Enterobacter cloacae ATCC 23355, Pseudomonas aeruginosa ATCC 27853, and Staphylococcus aureus ATCC 29213 with various combinations of cefepime or ceftazidime with tobramycin or ciprofloxacin using time-kill, checkerboard, and E test techniques. Time-kill testing was performed against each organism alone and in combinations at one-fourth times the MIC (1/4 x MIC) and 2 x MIC. With checkerboard tests, the same combinations were studied at concentrations ranging from 1/32 x to 4 x MIC. Standard definitions for synergy, indifference, and antagonism were utilized. E test strips were crossed at a 90 degree angle so the scales met at the MIC of each drug alone, and the fractional inhibitory concentrations index was calculated on the basis of the resultant zone on inhibition. All antimicrobial combinations demonstrated some degree of synergy against the test organisms, and antagonism was infrequent. Agreement with time-kill testing ranged from 44 to 88% and 63 to 75% by the checkerboard and E test synergy methods, respectively. Despite each of these methods utilizing different conditions and endpoints, there was frequent agreement among the methods. Further comparisons of the E test synergy technique with the checkerboard and time-kill methods are warranted.
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            Synergy tests by E test and checkerboard methods of antimicrobial combinations against Brucella melitensis.

            Yasemin Zer, G Orhan, Iclal Balci … (2004)
            Two different synergy testing methods, the checkerboard and the E test methods, were used to compare the in vitro efficacies of various antimicrobial combinations against 16 Brucella melitensis strains isolated from blood cultures. The rate of agreement of the E test and checkerboard methods was found to be 55%. The most concordant results were found for the streptomycin-doxycycline combination in 12 (75%) tests, in which four strains showed synergistic activity by E test and antagonistic activity by the checkerboard method and in which one strain showed antagonistic activity by both methods. Even though each of these methods uses different conditions and endpoints, the results of both methods frequently agreed.
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              Comparison of Etest, chequerboard dilution and time-kill studies for the detection of synergy or antagonism between antifungal agents tested against Candida species.

              A Messer, Michael E. Klepser, M Pfaller … (2002)
              Currently, there is considerable debate regarding the best in vitro method for testing antifungal combinations against Candida spp. In this study, we compared the results obtained by chequerboard dilution, time-kill studies and Etest for several antifungal combinations against Candida spp. Three Candida albicans isolates (fluconazole MICs of 1.0, 32 and >256 mg/L) and three non-albicans Candida isolates (C. glabrata, C. tropicalis and C. krusei) were tested in RPMI 1640 medium. By chequerboard testing, the majority of antifungal combinations were found to be indifferent. Notably, antagonism was identified by time-kill studies and by Etest for combinations of amphotericin B-fluconazole, but it was not detected by the chequerboard method. Pre-exposure of isolates to fluconazole did not affect results of the Etest or chequerboard method, but it did increase the frequency of antagonism noted by time-kill methods. This study indicates that chequerboard dilution testing in RPMI medium may not reliably detect the attenuation of amphotericin B activity. Of the three methods, Etest was the simplest to use and yielded reproducible results for testing antifungal combinations.
              Article 0 comments Cited 49 times – based on 0 reviews     Review now
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                Author and article information

                Journal
                Journal ID (nlm-ta): Braz J Microbiol
                Journal ID (iso-abbrev): Braz. J. Microbiol
                Journal ID (publisher-id): bjm
                Journal ID (pmc): bjm
                Title: Brazilian Journal of Microbiology
                Publisher: Sociedade Brasileira de Microbiologia
                ISSN (Print): 1517-8382
                ISSN (Electronic): 1678-4405
                Publication date (Print): Apr-Jun 2008
                Publication date (Electronic): 1 June 2008
                Volume: 39
                Issue: 2
                Pages: 233-237
                Affiliations
                [1 ]Refik Saydam National Hygiene Center, Department of Communicable Diseases Research
                [2 ]Gazi University Faculty of Medicine, Department of Infections Diseases
                Author notes
                *Corresponding Author. Mailing address: Refik Saydam National Hygiene center, Communicable Diseases Research, Cemal Gursel Street no:18 Sihhiye, Ankara. Tel.: +90-312-458 21 69. E-mail: selcuk.kilic@ 123456rshm.gov.tr
                Article
                Publisher ID: S1517-83822008000200006
                DOI: 10.1590/S1517-83822008000200006
                PMC ID: 3768412
                PubMed ID: 24031207
                SO-VID: f5aff685-d76e-43d3-a8b4-5b2649de2fe8
                Copyright © © Sociedade Brasileira de Microbiologia
                License:

                All the content of the journal, except where otherwise noted, is licensed under a Creative Commons License

                History
                Date received : 21 June 2007
                Date revision received : 22 September 2007
                Date accepted : 20 January 2008
                Categories
                Subject: Medical Microbiology
                Subject: Short Communication

                Keywords: brucella melitensis,synergism.,antibiotics
                Data availability:
                Keywords: brucella melitensis, synergism., antibiotics

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