ABSTRACT
Due to the Fourier-Transform Infrared Spectroscopy (FT-IR) of strain specific traits demonstrated to be a suitable and efficient method for diagnostic and epidemiological determinations for the yeasts Candida albicans, Exophiala dermatitidis and the chlorophylless algae of the genus Prototheca. FT-IR leads in a rapid and economical way to reproducible results according to the spectral differences of intact cells (IR-fingerprints). Different genera, species and sub-species respectively, different strains can be recognized and grouped into different clusters and subclusters. The FT-IR analysis of Candida albicans isolates (n = 150) of 22 newborns-at-risk of an intensive care unit showed, that 86% of the children were colonised with several (2-4) different strains in the oral cavities and faeces. Stationary cross-infections could definitely be determined. Exophiala dermatitidis isolates (n = 31), mostly isolated repetitively within a period of 3 years from sputa of patients suffering from cystic fibrosis could be characterized and grouped patient-specifically over the total sampling period. Of 6 from 8 patients (75%) their individual strains remain the same and could be tracked over the three years. Cross-infections during the stationary treatment could be clearly identified by FT-IR. The Prototheca isolate (n = 43) from live-stock and farm environment showed clear distinguishable clusters differentiating the species P. wickerhamii, P. zopfii and P. stagnora. In addition, the biotypes of P. zopfii could be distinguished, especially the subclusters of variants II and III. It could be demonstrated, that FT-IR is suitable for the routine identification and differentiation of yeasts and algae. However, in spite of the gain of knowledge by using FT-IR for the characterization of microorganisms, the conventional phenotyping and/or genetic analysis of yeast or algae strains cannot be replaced completely. For a final taxonomic classification a combination of conventional methods on FT-IR together with more sophisticated molecular genetic procedures is necessary.
Subject(s)
Animals, Domestic , Candida albicans/classification , Exophiala/classification , Infections/veterinary , Mycoses/microbiology , Prototheca/classification , Animals , Candidiasis/diagnosis , Candidiasis/epidemiology , Candidiasis/microbiology , Child, Preschool , Humans , Infant , Infant, Newborn , Infections/microbiology , Mycoses/diagnosis , Mycoses/epidemiology , Spectroscopy, Fourier Transform InfraredABSTRACT
The in-vitro activity of azithromycin, clarithromycin, erythromycin, josamycin, midekamycin, roxithromycin and clindamycin against 674 Gram-negative and Gram-positive clinical isolates, including methicillin-resistant Staphylococcus aureus, was determined by agar dilution, microdilution and agar diffusion with Mueller-Hinton medium according to the Deutsches Institut für Normung (DIN) 58940 guidelines. The results obtained by regression analysis and the error-rate-bounded method of Metzler-DeHaan indicate that common interpretative criteria (breakpoints) for test discs may be assigned to susceptible/resistant Gram-positive strains for all antibiotics tested. The following tentative DIN values are suggested for 15 microg macrolide discs: for susceptible Gram-positive and Gram-negative strains, an inhibition zone diameter (IZD) of > or = 26 mm at a corresponding MIC of < or = 2 mg/L; for resistant Gram-positive strains, an IZD of < or = 21 mm; for resistant Gram-negative strains, an IZD of < or = 19 mm at a corresponding MIC of > or = 8 mg/L. For Haemophilus influenzae only, breakpoints for azithromycin are suggested with IZDs of > or = 21 mm for susceptible and < or = 18 mm for resistant.
Subject(s)
Anti-Bacterial Agents/pharmacology , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Microbial Sensitivity Tests , Dose-Response Relationship, Drug , Guidelines as Topic , Haemophilus influenzae/drug effects , Humans , Lincosamides , Macrolides/pharmacology , Methicillin Resistance , Staphylococcus aureus/drug effectsABSTRACT
In bacteriology, the Etest has a broad field of application in bacteriology and is recently available for the antimycotics fluconazole and itraconazole. By means of the presence of gradient concentrations of the active substance on the carrier material, it is possible to obtain reproducible MICs of the antimycotic substances. The results of susceptibility testing of 326 clinical yeast isolates with the Etest were compared to those MICs obtained by microdilution and agar dilution. A 100% concordance of the MIC markers (mode-, MIC50- and MIC90-value, standard deviation of the mean log2-MIC-dilution steps) was given when compared by a +/- 1 MIC-dilution step range with microdilution and by +/- 2 MIC-dilution steps with agar dilution; species dependent all strains were within 2 x standard deviation of the individual MIC-mean of the species. By comparison of the individual MIC-values maximum differences of +/- 6 MIC-dilution steps were obtained, where 70% of all results were within +/- 2 MIC-dilution steps, and more than 92% of all strains were within +/- 3 MIC-dilution steps. The Pearson's correlation coefficients show a good agreement of the Etest with microdilution (r = 0.92) resp., agar dilution (r = 0.88) demonstrate, however, clearly insufficient correlations (r < 0.65) to the reference methods, for species with difficult to read Etest inhibition zones (e.g., Candida glabrata, Candida krusei, Candida parapsilosis). The differences between the proposed test methods recommended by the NCCLS and the working group "Clinical Mycology" of the German Speaking Mycological Society (AG-KMYK) are tabled.
Subject(s)
Antifungal Agents/pharmacology , Candida/drug effects , Fluconazole/pharmacology , Microbial Sensitivity Tests/methods , Yeasts/drug effects , Agar , Candida/isolation & purification , Humans , Microchemistry , Mycoses/microbiology , Yeasts/isolation & purificationABSTRACT
Four commercially available in vitro test systems (Candifast, E-test, Mycototal and spiral-gradient end point method), agar diffusion with 25-micrograms fluconazole paper test discs and 15-micrograms test tablets and agar dilution were compared with the microbroth dilution method for fluconazole susceptibility testing of 145 clinical isolates. In addition, the culture media provided or recommended by the manufacturers of the test systems were compared with high-resolution (HR) antifungal test medium. With all currently available culture media, growth problems (inhibition or delayed growth of the clinical isolates) occurred with solid or semisolid media. With minor improvements, HR medium demonstrated the most reproducible and comparable results (supplementation with asparagine and deletion of sodium hydrogen carbonate). The best correlation with microdilution was obtained by the agar dilution method (> 95% concordance) followed by the spiral-gradient end point method (85%), Candifast (83%), Mycototal (81%) and the E-test (78%). Regression analysis demonstrated good correlation between agar diffusion and micro-/agar dilution (r > 0.9).
Subject(s)
Antifungal Agents/pharmacology , Fluconazole/pharmacology , Fungi/drug effects , Candida/drug effects , Candida/isolation & purification , Cryptococcus/drug effects , Cryptococcus/isolation & purification , Culture Media , Fungi/growth & development , Fungi/isolation & purification , Geotrichum/drug effects , Geotrichum/isolation & purification , Humans , Microbial Sensitivity Tests/methods , Regression Analysis , Rhodotorula/drug effects , Rhodotorula/isolation & purification , Trichosporon/drug effects , Trichosporon/isolation & purificationABSTRACT
Four commercially available in vitro test systems (Candifast, E-test, Mycototal, Spiral-Gradient Endpoint Method), agardiffusion with 25 micrograms fluconazole paper test discs and 15 micrograms test tablets, and agardilution were compared to the microbroth dilution method by fluconazole susceptibility testing of 145 clinical isolates. In addition, the culture media provided or recommended by the manufacturers of the test systems were compared to the high resolution (HR) antifungal test medium. With all currently available culture media growth problems (inhibition or delayed growth of the clinical isolates) occurred with solid or semi-solid media. With minor improvements, HR medium demonstrated the most reproducible and comparable results (supplementation with asparagine and deletion of sodium hydrogen carbonate). Best correlation to microdilution was obtained by the agardilution method > (95% concordance) followed by the spiral gradient endpoint method (85%), Candifast (83%), Mycototal (81%) and the E-test (78%). Regression analysis demonstrated good correlation between agardiffusion and micro-/agardilution(r > 0.9).
