agar and Mycoses

Topics: Actinomyces; Agar; Arthrodermataceae; Candida; Cryptococcus; Cycloheximide; Fungi; Glucose; Mycoses; Nocardia; Staining and Labeling; Urease

In the present report, we describe an unusual case of mixed infection of Candida albicans and Talaromyces marneffei in the oral cavity and oropharynx with cutaneous involvement. On the CHROMagar Candida plate, green colonies (identified as C. albicans) and tiny violet colonies (identified as T. marneffei) grew from the throat swab after incubation for 96 hours. 10 clinical isolates of T. marneffei were used to verify their color production on CHROMagar Candida. All colonies were violet on the fourth, seventh and ninth day incubated at 37 °C. T. marneffei appears violet on the CHROMagar Candida plate, but it may be easily ignored because of its slow growth and small colony size, especially after incubation for 48 hours. Therefore, when using CHROMagar Candida plate to detect specimens in AIDS patients, special attention must be paid to detect non-yeasts such as T. marneffei for up to 96 hours.

Topics: Agar; AIDS-Related Opportunistic Infections; Candida albicans; Coinfection; Culture Media; Humans; Male; Middle Aged; Mouth; Mycological Typing Techniques; Mycoses; Oropharynx; Talaromyces; Time Factors

We developed a novel culture medium, referred to FastFung medium as suitable for the culture of clinical fungi, including fastidious ones, for both research and diagnostic studies. It is based on Schædler agar supplemented with many essential components for the growth of fastidious fungi. It also contains selective antibacterial agents for the inhibition of contaminant bacteria growth. In this preliminary study, the FastFung medium was compared to the gold standard Sabouraud medium for 98 fungal and 20 bacterial strains. The fungal strain positive culture rate was 100% vs. 95% and the bacterial strain inhibition was 100% vs. 20%, for the FastFung and Sabouraud media, respectively. When compared to the Sabouraud medium on 120 clinical samples, the FastFung medium displayed both a higher fungal colonies count, and a lower culture contamination rate. Storage at 4 °C for 4 weeks did not alter the FastFung culture medium performances for the six isolates of Candida, Cryptococcus, and Penicillium tested. These encouraging results suggest future development of using the FastFung medium in clinical mycology and in mycobiome characterization. Further prospective evaluation aiming at assessing whether implementing the FastFung medium in the routine workflow simplifies and strengthen fungal isolation capacities in the clinical laboratory is warranted.

Topics: Agar; Bacteria; Candida; Clinical Laboratory Techniques; Cryptococcus; Culture Media; Fungi; Genes, rRNA; Malassezia; Mycobiome; Mycology; Mycoses; Penicillium

Phaeoacremonium parasiticum is an environmental dematiaceous mold rarely associated with human infections. We present here 2 cases of P. parasiticum invasive infections, including the first report of P. parasiticum respiratory tract infection, and 1 case of airway colonization, which all 3 strains of P. parasiticum were identified using agar block smear and ITS and β-tubulin gene sequencing. All 3 isolates grew initially as white to creamy, yeast-like colonies. After 21 days of incubation at 25 °C, 1 isolate remained light brown, atypical of P. parasiticum. Microscopic examination of agar block smear preparations of all 3 isolates showed thick-walled, medium brown conidiophores that were branched and slightly swollen at the base. The sequences of the ITS and β-tubulin genes of the 3 isolates were identical to those of P. parasiticum. Cases of P. parasiticum infections should be confirmed by a polyphasic approach using morphologic characterization and ITS and β-tubulin gene sequencing.

Topics: Adult; Agar; Aged; Ascomycota; Culture Media; DNA, Fungal; DNA, Ribosomal Spacer; Humans; Male; Microscopy; Molecular Sequence Data; Mycoses; Phylogeny; Respiratory Tract Infections; Sequence Analysis, DNA; Tubulin

Clinical specimens cultured on two selective fungal media, inhibitory mold agar (IMA) and Sabouraud dextrose agar (SDA), were compared with respect to recovery of fungi. Of the 840 fungal isolates recovered, 69.3% grew on both IMA and SDA; 24.9% grew only on IMA; and 5.8% grew only on SDA, showing that IMA is superior (P=0.003).

Topics: Agar; Culture Media; Fungi; Glucose; Humans; Mycology; Mycoses; Sensitivity and Specificity

The objective of this prospective study was to assess the prevalence of Exophiala dermatitidis in respiratory secretions of patients with cystic fibrosis (CF) and to identify risk factors for its presence. The results of all cultures performed over a 2-year period in non lung-transplant patients in our CF clinic were included in the study. Samples consisted of sputum (whenever possible) or deep pharyngeal aspirate after a session of physiotherapy. Specimens were inoculated onto Sabouraud gentamicin-chloramphenicol agar (SGCA) medium (Becton-Dickinson) and incubated at 35°C for 2 days and then at ambient temperature (15-25°C) for 3 weeks. The whole study group included 154 patients (mean age ± SD: 18.5 y ± 11.69). E. dermatitidis was isolated from 58 specimens (2.8%) of nine patients (5.8%) out of total of 2065 cultures prepared during the study period. All E. dermatitidis culture-positive patients were pancreatic insufficient and ≥12 y of age. Almost all (8/9) were homozygous for the F508 del mutation. Aspergillus fumigatus colonization and genotype seemed to be predisposing factors. No other significant characteristic was identified in this group, either in terms of predominant bacterial pathogen or treatment. A distinct comparative study performed over 3 months in our laboratory revealed that the use of SGCA yielded identical isolation rates of E. dermatitidis as erythritol-chloramphenicol agar (ECA).

Topics: Adolescent; Adult; Agar; Child; Child, Preschool; Culture Media; Cystic Fibrosis; Exophiala; Female; Humans; Infant; Lung Diseases, Fungal; Male; Microbiological Techniques; Middle Aged; Mycoses; Prevalence; Risk Factors; Sputum; Young Adult

Incorporation of gallium (III) nitrate into unsupplemented Sabouraud Dextrose Agar to a final concentration of 512 mg/l (2 mM) suppressed bacterial growth, of the following genera Escherichia, Enterococcus, Klebsiella, Listeria, Pseudomonas and Staphylococcus, In contrast growth of Burkholderia cenocepacia, and yeast and filamentous fungi was not affected. Supplementation of selective mycological media with gallium (III) may aid in the selectivity of such media, particularly where clinical specimens are heavily contaminated with bacterial co-habitants and where antibiotic resistance in such bacterial flora may render antibiotic supplements ineffective.

Topics: Agar; Bacteria; Culture Media; Drug Resistance, Bacterial; Gallium; Humans; Iron; Microbiological Techniques; Mitosporic Fungi; Mycoses; Sensitivity and Specificity; Trace Elements

Chromogenic Candida Agar is a novel differential culture medium that is claimed to facilitate isolation and identification of Candida albicans, Candida tropicalis and Candida krusei. The performance of this medium was evaluated for presumptive identification of 521 yeast strains, representing 23 different species, for detection of specimens containing yeast mixtures, and for direct isolation of yeast from blood cultures. All yeasts grew well on the medium following a 48-h incubation period at 37 degrees C, and distinctive colonies were produced by C. albicans, C. tropicalis, C. krusei, Candida guilliermondii, Saccharomyces cerevisiae, Trichosporon mucoides and Geotrichum capitatum. The sensitivity and specificity of the medium exceeded 99.4% for each of these species. The medium provided some indication of the presence of Candida dubliniensis and Candida pulcherrima, and allowed the identification of polyfungal samples in 89.4% of the yeast mixtures. Finally, direct isolation on the medium from blood cultures that were positive for yeast according to Gram's stain (n = 42) showed that the expected colour and morphology of each species were not altered in the presence of blood.

Topics: Agar; Blood; Chromogenic Compounds; Culture Media; Humans; Mycological Typing Techniques; Mycoses; Sensitivity and Specificity; Species Specificity; Yeasts

Data on clinical isolates of Kodamaea (Pichia) ohmeri, an emerging fungal pathogen, are scarce. Over the past 5 years, we identified yeast isolates from nine patients with fungemia as K. ohmeri by using the API 20C system. Here, we reanalyzed these isolates first by sequencing the internal transcribed spacer 2 (ITS2) regions and then by growing the isolates on CHROMagar Candida medium and subjecting them to pulsed-field gel electrophoresis (PFGE). Based on their ITS2 sequences, six of the nine isolates were confirmed as K. ohmeri, while the others were identified as Candida haemulonii (n = 2) and Candida parapsilosis (n = 1). PFGE karyotyping of the K. ohmeri isolates revealed similar major bands, and their colonies showed a characteristic color change from pink to blue when grown on CHROMagar Candida medium for more than 48 h. For K. ohmeri, the ranges of MICs of fluconazole, voriconazole, caspofungin, and micafungin were 2 to 32 mug/ml, 0.03 to 0.5 mug/ml, 0.125 to 0.25 mug/ml, and 0.03 to 0.06 mug/ml, respectively. Restriction endonuclease analysis of genomic NotI-digested DNA (REAG-N) from isolates from different patients produced unique patterns, suggesting that the fungemia had occurred sporadically. This study determined that ITS2 sequence data, PFGE karyotypes, and CHROMagar Candida chromogenic culture medium are reliable diagnostic tools for identifying K. ohmeri while REAG-N is useful for genotyping the clinical isolates of K. ohmeri.

Topics: Agar; Antifungal Agents; Chromogenic Compounds; Culture Media; DNA, Ribosomal Spacer; Electrophoresis, Gel, Pulsed-Field; Fungemia; Hospitals, University; Humans; Microbial Sensitivity Tests; Mycological Typing Techniques; Mycoses; Pichia; Saccharomycetales; Sequence Analysis, DNA

To establish the best method for boric acid susceptibility testing, we compared two agar dilution methods (high and low inoculum) and a standard broth microdilution method (from the National Commitee for Clinical Laboratory Standards document NCCLS M-27A). Saccharomyces cerevisiae (37) and non-C. albicans Candida (39) isolates, as well as one isolate of Trichosporon sp., were included. All were isolated from female workers with vulvovaginitis. Good agreement within a fourfold dilution range was found between the three methods, and only the broth microdilution method versus the agar dilution method with high inoculum showed significant discrepancies. Reading results was easier with the broth microdilution method than with the agar dilution methods because of partial growth inhibition in the latter. In conclusion, broth microdilution is a suitable method for testing yeast susceptibility to boric acid.

Topics: Agar; Anti-Infective Agents, Local; Antifungal Agents; Boric Acids; Candida; Candidiasis, Vulvovaginal; Culture Media; Female; Humans; Microbial Sensitivity Tests; Mycoses; Saccharomyces cerevisiae; Trichosporon; Vulvovaginitis

Candida ID is a new chromogenic medium for the identification of yeasts from clinical specimens. C. albicans produces blue pigmentation, whereas pink pigmentation is produced by C. tropicalis, C lusitaniae, C. guilliermondii and C. kefyr; other Candida species appear white. In this study, 240 clinical samples (throat swabs and stool samples) from haematology patients were inoculated on to Candida ID and Sabouraud-chloramphenicol agar in parallel, yielding a total of 105 yeasts; the media had overall detection rates of 85.7% and 86.7% respectively. The sensitivity of Candida ID for identification of C. albicans by blue pigmentation was 52.9% at 24 h and 94.1% at 48 h. Specificity of the blue pigmentation was 100% at 48 h. Two strains of C. tropicalis were identified, one produced pink pigmentation at 72 h, the other strain did not produce any pigmentation after 5 days. Candida ID was superior in detecting mixtures of yeasts compared with Sabouraud-chloramphenicol agar. Candida ID is a suitable primary isolation medium for yeasts from clinical specimens, providing rapid direct identification of C. albicans and enhanced detection of mixtures.

Topics: Agar; Chloramphenicol; Chromogenic Compounds; Culture Media; Hospital Units; Humans; Mycological Typing Techniques; Mycoses; Population Surveillance; Sensitivity and Specificity; Time Factors; United Kingdom; Yeasts

This report presents a semisolid agar antifungal susceptibility (SAAS) method for the rapid susceptibility screening of yeasts and molds. The reproducibility and accuracy of the SAAS method were assessed by comparing the MICs of amphotericin B and fluconazole obtained for 10 candidate quality control (QC) American Type Culture Collection yeast strains in >/=15 replicates with those found by six independent laboratories using the National Committee for Clinical Laboratory Standards (NCCLS) M27-P broth macrodilution method (M. A. Pfaller et al., J. Clin. Microbiol. 33:1104-1107, 1995). Overall, 96% of MICs for both drugs fell within 1 log(2) dilution of the modal MIC for each strain. The MICs for amphotericin B showed 99% agreement with the NCCLS proposed QC ranges within 1 log(2) dilution. Likewise, the MICs for fluconazole at >/=75% growth reduction showed 99% agreement for seven strains. Three strains, Candida albicans ATCC 24333 and ATCC 76615 and Candida tropicalis ATCC 750, showed a less sharp fluconazole endpoint at >/=75% growth reduction, but at >50% growth reduction, the agreement was 98% within 1 log(2) dilution of the proposed range. The MIC agreement within the proposed range for the suggested QC strains Candida parapsilosis ATCC 22019 and Candida krusei ATCC 6258 was 100% for fluconazole and 100% within 1 log(2) dilution of the proposed range for amphotericin B. The SAAS method demonstrated the susceptibility or resistance of 25 clinical isolates of filamentous fungi such as Aspergillus fumigatus to amphotericin B, itraconazole, and fluconazole, usually within 48 h. Although the results are preliminary, this SAAS method is promising as a rapid and cost-effective screen and is worthy of concerted investigation.

Topics: Agar; Antifungal Agents; Culture Media; Evaluation Studies as Topic; Fungi; Humans; Microbial Sensitivity Tests; Mycoses; Reproducibility of Results; Sensitivity and Specificity

Enriched broth medium is routinely used as a supplement for agar plate culture of cerebrospinal fluid (CSF). To assess the clinical utility of broth cultures, 151 consecutive CSF bacterial and fungal isolates obtained from 91 patients were retrospectively reviewed for the effect of results on treatment. Treatment decisions associated with individual CSF specimens for which isolates were recovered from thioglycollate broth only were compared with the treatment decisions associated with CSF specimens for which isolates were recovered by agar plate culture. Treatment was defined as initiation of or change in antimicrobial therapy based on the reporting of CSF culture isolates. Thirty-six (24%) of the 151 isolates were recovered in broth only. Three (8%) of these 36 isolates (from 34 patients) resulted in treatment with antimicrobial agents; however, 2 of the 3 treated isolates (Candida tropicalis, Proteus mirabilis) were recovered from a second CSF specimen in agar plate culture within 24 hours. Thus, only a single isolate (3%; Staphylococcus epidermidis) was treated based solely on a positive broth culture result. In contrast, 60 (52%) of the 115 isolates recovered in agar plate culture from 23 (40%) of 57 patients were treated (staphylococci, 28; gram-negative bacilli, 14; Cryptococcus neoformans, 10; Streptococcus pneumoniae, 3; Streptococcus sanguis, 1; other, 4). We conclude that treatment with antimicrobial agents based on isolates recovered from CSF specimens in broth culture alone is infrequent and infer from the data that the use of CSF broth cultures contributes little to treatment decisions.

Topics: Adolescent; Adult; Agar; Aged; Aged, 80 and over; Anti-Bacterial Agents; Anti-Infective Agents; Bacteria; Bacterial Infections; Cerebrospinal Fluid; Culture Media; Female; Fungi; Humans; Infant; Male; Microbiological Techniques; Middle Aged; Mycoses; Retrospective Studies

Two chromogenic media, Albicans ID and CHROMagar Candida agar plates, were compared with a reference medium, Sabouraud-chloramphenicol agar, and standard methods for the identification of yeast species. This study involved 951 clinical specimens. The detection rates for the two chromogenic media for polymicrobial specimens were 20% higher than that for the Sabouraud-chloramphenicol agar plates. The rates of identification of Candida albicans for Albicans ID and CHROMagar Candida agar plates were, respectively, 37.0 and 6.0% after 24 h of incubation and 93.6 and 92.2% after 72 h of incubation, with specificities of 99.8 and 100%. Furthermore, CHROMagar Candida plates identified 13 of 14 Candida tropicalis and 9 of 12 Candida krusei strains after 48 h of incubation.

Topics: Agar; Candida; Candida albicans; Candidiasis; Chromogenic Compounds; Culture Media; Evaluation Studies as Topic; Humans; Mycology; Mycoses; Yeasts

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.

Topics: Agar; Antifungal Agents; Candida; Fluconazole; Humans; Microbial Sensitivity Tests; Microchemistry; Mycoses; Yeasts

The updated Vitek Yeast Biochemical Card (YBC) was compared with the API 20C by using 409 germ tube-negative yeasts and Geotrichum spp. that were either clinical or proficiency sample isolates. The API 20C was the reference standard. The 409 isolates represented nine genera and 21 species. Morphology agars were inoculated and interpreted for each isolate. The API 20C identified 406 isolates (99.3%), while the Vitek YBC identified 367 (89.7%). Both systems identified the majority of yeasts after 24 h of incubation--73.4% were identified by the API 20C and 77.4% were identified by the Vitek YBC. The Vitek 24-h reading had some incorrect identifications. These included 14 isolates of Candida tropicalis that were identified as Candida parapsilosis (91 to 97% reliability) and 3 isolates of Candida krusei that were called Blastoschizomyces capitatus (Geotrichum capitatum), Candida rugosa, and Candida zeylanoides. In total, the Vitek YBC misidentified 30 isolates, while the API 20C misidentified 3 isolates. In addition, results for 14 isolates with the Vitek YBC were listed under the category "no identification." Morphology agars were required for identification with 89 isolates (21.9%) when the API 20C was used and with 50 isolates (12.6%) when the Vitek YBC was used. Apart from the price of the Vitek instrument, the API 20C costs $1.28 more per test than the Vitek YBC. Overall, the updated Vitek YBC compares favorably with the API 20C in the identification of common yeasts such as Torulopsis glabrata, C. parapsilosis, and Cryptococcus neoformans. However, problems were encountered with the Vitek system in the identification of C. tropicalis, C. krusei, Trichosporon spp., and some Cryptococcus spp. The routine use of morphology agars with either method is recommended.

Topics: Agar; Candidiasis; Costs and Cost Analysis; Evaluation Studies as Topic; Fungemia; Fungi; Humans; Mycology; Mycoses; Reproducibility of Results

Annellated conidiogenous cells of both the mononematous (Scedosporium state) and synnematous (Graphium state) conidiophores develop in the conidial state of Petriellidium (= Allescheria) boydii, Scedosporium (= Monosporium) apiospermum. Conidiogenesis in this species is characterized by percurrent proliferation of the conidiogenous cells and the conidia are holoblastic and annellidic.

Topics: Adult; Agar; Ascomycota; Cell Wall; Humans; Keratitis; Lung Diseases, Fungal; Male; Mycoses; Spores, Fungal

Motility and various biochemical activities of isolates of bacteria and yeasts were tested on undivided agar plates by using a simple, manually operated multipoint inoculation apparatus that allowed the analysis of 25 isolates per 9-cm-diameter petri plate. Fermentation of all 17 carbohydrates tested as well as 13 other biochemical activities commonly used for identification of bacteria were readily demonstrated by the multipoint inoculation plate method, and the results agreed very well with those of conventional tube tests. In addition to speedy inoculation and low cost of materials, the multipoint inoculation plate method offers several other advantages when compared with conventional tube tests or with some of the manufactured test kits currently available for recognizing members of the family Enterobacteriaceae.

Topics: Agar; Bacteria; Bacterial Infections; Bacterial Physiological Phenomena; Enterobacteriaceae; Humans; Microbiological Techniques; Mycoses; Yeasts

Topics: Agar; Blood; Carbohydrate Metabolism; Eukaryota; Glucose; Humans; Mycoses; Species Specificity; Spores; Temperature; Venezuela; Yeasts

Topics: Agar; Fungi; Mycoses; Precipitin Tests; Soil Microbiology

Topics: Adolescent; Adult; Agar; Aged; Anti-Bacterial Agents; Antineoplastic Agents; Autopsy; Belgium; Candida; Child; Cortisone; Digestive System; Female; Fungi; Humans; Male; Middle Aged; Mycoses; Neoplasms; Respiratory System; Sucrose

Topics: Agar; Bacillus; Culture Media; Feces; Food Microbiology; Fungi; Gentamicins; Glucose; Humans; Mycoses; Oxytetracycline; Saccharomyces; Soil Microbiology; Suppuration; Yeasts

Topics: Agar; Humans; Mycoses; Precipitin Tests

Topics: Agar; Diagnosis, Differential; Female; Humans; Methods; Mycoses; Nails; Vaginal Smears; Yeasts

Topics: Agar; Amylases; Animals; Caseins; Cattle; Fats; Fungi; Humans; Lipase; Milk; Mycoses; Oryza; Peptide Hydrolases; Starch

Topics: Adult; Agar; Anti-Bacterial Agents; Corneal Ulcer; Cortisone; Diagnosis, Differential; Female; Humans; Mycoses; Surgical Procedures, Operative; Yeasts

Topics: Agar; Anti-Infective Agents, Local; Biological Assay; Biological Products; Candida; Infection Control; Infertility; Mercury Compounds; Mycoses; Research; Staphylococcus; Thimerosal

Topics: Agar; Cycloheximide; Dermatology; Humans; Mycoses

Topics: Agar; Antibodies; Fungi; Humans; Mycobacterium; Mycoses; Sarcoidosis