agar and Candidiasis

The incidence of invasive candidiasis has increased dramatically over the last decades due to a larger number of patients at risk. The diagnosis remains difficult as the clinical presentation is not specific and the biological diagnosis usually takes several days to become positive. We propose in this work through a prospective study to evaluate the contribution of a chromogenic medium CHROMagar(®) (Becton-Dickinson) in the mycological diagnosis of Candida. We selected 680 samples from patients hospitalized in the intensive care unit for epidemiological surveillance over a period of 11 weeks. We treated samples by culture on Sabouraud and on CHROMagar(®). The species identification was performed by chlamydosporulation test and carbohydrate assimilation tests. We found that the CHROMagar(®)Candida evaluated in our work was a valuable tool in the primary culture in differentiating the most frequently isolated yeast species and in better detection of mixed cultures.

Topics: Agar; Axilla; Candida; Candidiasis; Chromogenic Compounds; Cross Infection; Culture Media; Fungal Proteins; Hexosaminidases; Hospitals, Military; Humans; Inpatients; Intensive Care Units; Monophenol Monooxygenase; Mouth; Nasal Cavity; Population Surveillance; Predictive Value of Tests; Prospective Studies; Rectum; Sensitivity and Specificity; Species Specificity; Tunisia

Candidemia and vaginitis are the most common types of candidiasis mostly caused by Candida albicans species. C. albicans has several genotypes and the potential ability to form different phenotype colonies on specific media. This study aimed to evaluate the genotype distribution of blood and vaginal C. albicans isolates and phenotype characteristics on Spider and yeast peptone dextrose agar medium.. A total of 40 clinical Candida albicans isolates comprising vagina (20) and blood (20) were used. ABC typing using CA-INT-R and CA-INT-L primers was performed to span the transposable group I intron of the 25S rDNA gene. For colony phenotypic characteristics, the Spider and YPDA media were used.. Among the blood and vaginal isolates, genotype A (12/60%) and genotype C (10/50%) were the most common types, respectively. The highest phenotype shape frequency of the colonies in blood and vaginal samples was the ring and the lowest was the hat/ring. The dominant color phenotype in blood and vaginal samples was gray. There was a significant relationship between genotype and phenotype forms in the blood sample on YPDA medium (p = 0.02). In the Spider medium, there were no significant differences between genotypes and phenotypes.. In this study, genotype A and genotype C were predominant in blood and vaginal samples, respectively. In both groups, YPD agar medium demonstrated the most variety of phenotypes that was related to genotypes A and C. The variety of phenotypes in both groups was the same in genotypes A and C on the Spider medium.

Topics: Agar; Animals; Candida; Candida albicans; Candidiasis; Genotype; Phenotype

Skin colonization by the emerging pathogen Candida auris is common in outbreaks within medical settings. Culture-based screening of patients is an effective management strategy to control the pathogen, and the newly developed CHROMagar™ Candida Plus medium is claimed to enable the presumptive identification of C. auris. Here, we evaluated the use of this medium with 63 C. auris strains comprising its four well-established clades, as well as genetically related comparators, including species from the Metschnikowia clade. The colors and halos of both confluent growth and discrete colonies of all the tested strains were compared. It was found that on CHROMagar™ Candida Plus, C. auris formed characteristic white colonies with blue-green halos that were more evident after 72 hr of incubation at 35°C than after 48 hr. However, distinguishing between closely related species such as Candida haemulonii, Candida pseudohaemulonii, and Candida duobushaemulonii required the consideration of parameters other than color, including colony size and growth ability at 35°C. In conclusion, the novel chromogenic medium CHROMagar™ Candida Plus constitutes an easy screening tool for C. auris.

Topics: Agar; Antifungal Agents; Candida; Candida auris; Candidiasis; Humans

Candidal vulvovaginitis is one of the most common genital infections that different types of diagnosis are essential for a proper treatment plan. IUD is one of the most influential and long-lasting methods of contraception that can be associated with vaginal candidiasis. This study was performed to investigate the prevalence of Candida species before and three months after IUD placement in patients referred to health centers. Also, a comparison of copper IUDs and hormonal IUDs was evaluated to consider the prevalence of Candida species in cervicovaginal smears. In this regard, cervicovaginal swabs were prepared from 160 women applying for IUDs who did not show signs of vaginal infection during the vaginal examination. These people were divided into two groups of 80 cases. The first group received copper IUDs (NT Cu380, Mona Lisa®, Canada), and the second group received hormonal IUDs (Mirena, USA). They had not used antibiotics or antifungal drugs at least two weeks before and three months after IUD placement. The provided Samples were cultured in a Saburo dextrose agar medium. The milky yeast colony was transferred to chromium agar culture medium, and fungal species were differentiated by dyeing. P <0.05 was considered significant. Three months after IUD insertion, 29.57% of people who received a copper IUD were diagnosed with candidiasis. Also, different species of Candida were observed in 22.95% of people who received hormonal IUD. Because Candida albicans is found in the vaginal microflora of 30 to 80% of asymptomatic women, the decision to treat asymptomatic cases requires further study and testing. The use of Candida chromium agar differential culture medium is easy, reproducible, and cost-effective; however, in cases such as recurrent or complicated vulvovaginal candidiasis where the accurate diagnosis is essential for successful treatment, the use of sensitive and precise molecular methods such as PCR is recommended. Finally, studies with wider dimensions and longer follow-up periods are suggested to confirm and complete the present study.

Topics: Agar; Candida; Candidiasis; Chromium; Female; Humans; Intrauterine Devices; Prevalence

Identification of the emerging multidrug-resistant yeast Candida auris is challenging. Here, we describe the role of the Mexico national reference laboratory Instituto de Diagnóstico y Referencia Epidemiológicos Dr. Manuel Martínez Báez (InDRE) and the Mexican national laboratory network in the identification of C. auris. Reference identification of six suspected isolates was done based on phenotypic and molecular laboratory methods, including growth in special media, evaluation of isolate micromorphology, and species-specific PCR and pan-fungal PCR and sequencing. The four C. auris isolates identified were able to grow on modified Sabouraud agar with 10% NaCl incubated at 42 °C. With one exception, isolates of C. auris were spherical to ovoid yeast-like cells and blastoconidia, with no hyphae or pseudohyphae on cornmeal agar. C. auris isolates were resistant to fluconazole. Species-specific and pan-fungal PCR confirmed isolates as C. auris. Sequence analysis revealed the presence of two different C. auris clades in Mexico, clade I (South Asia) and clade IV (South America).

Topics: Agar; Antifungal Agents; Candida; Candida auris; Candidiasis; Mexico; Microbial Sensitivity Tests

Candida auris is a serious nosocomial health risk, with widespread outbreaks in hospitals worldwide. Successful management of such outbreaks has depended upon intensive screening of patients to identify those that are colonized and the subsequent isolation or cohorting of affected patients to prevent onward transmission. Here we describe the evaluation of a novel chromogenic agar, CHROMagarTM Candida Plus, for the specific identification of Candida auris isolates from patient samples. Candida auris colonies on CHROMagarTM Candida Plus are pale cream with a distinctive blue halo that diffuses into the surrounding agar. Of over 50 different species of Candida and related genera that were cultured in parallel, only the vanishingly rare species Candida diddensiae gave a similar appearance. Moreover, both the rate of growth and number of colonies of C. auris recovered from swabs of pure and mixed Candida species were substantially increased on CHROMagarTM Candida Plus agar when compared with growth on the traditional mycological isolation medium, Sabouraud dextrose agar. Taken together, the present data suggest that CHROMagarTM Candida Plus agar is an excellent alternative to current conventional mycological media for the screening of patients who are potentially colonized/infected with Candida auris, can be reliably used to identify this emerging fungal pathogen, and should be tested in a clinical setting.. Candida auris is a novel pathogenic yeast that has been associated with large hospital outbreaks across several continents. Affected patients become colonized, predominantly on the skin, with large quantities of C. auris which they then shed into the hospital environment. Identification of C. auris is challenging using routine laboratory methods, and time consuming when patients are colonized with a mixture of different Candida species. Here we demonstrate that a novel chromogenic agar, CHROMagarTM Candida Plus, permits the rapid differentiation of C. auris from a wide range of other yeast species and is potentially ideally suited to screening of patients that are suspected of being colonized or infected with this medically important yeast.

Topics: Agar; Candida; Candidiasis; Culture Media; Humans; Microbiological Techniques; Saccharomycetales

The aim of this study was to compare the Candida bromcresol green (BCG) medium with the chromogenic (CHROM) Brilliance Candida agar and Sabouraud dextrose agar (SDA) media in regard to their capability of detecting Candida isolates from mono- or dual-species cultures. We prepared Candida isolates' suspensions to obtain mono-species (n = 18) or dual-species (n = 153) culture plates per each medium, and three readers independently observed 513 plates at 24-h, 48-h and 72-h incubation time. We scored reading results as correct, over or under detection compared to the expected species number(s). BCG showed significantly higher correct-detection and lower under-detection rates for all Candida species when observed by at least one reader. At 24-h reading, 12 mono-species cultures had correct (or over) detections in all media, whereas 106, 60 and 78 dual-species cultures had correct (or over) detections in BCG, CHROM or SDA, respectively. BCG provides the basis for an accurate laboratory diagnosis of Candida infections.

Topics: Agar; Candida; Candidiasis; Culture Media; Humans; Indicators and Reagents; Microbiological Techniques

The in vitro interactions of isavuconazole with colistin were evaluated against 15 clinical Candida auris isolates by a microdilution checkerboard technique based on the EUCAST reference method for antifungal susceptibility testing and by agar diffusion using isavuconazole gradient concentration strips with or without colistin incorporated RPMI agar. Interpretation of the checkerboard results was done by the fractional inhibitory concentration index and by response surface analysis based on the Bliss model. By checkerboard, combination was synergistic for 93% of the isolates when interpretation of the data was done by fractional inhibitory concentration index, and for 80% of the isolates by response surface analysis interpretation. By agar diffusion test, although all MICs in combination decreased compared to isavuconazole alone, only 13% of the isolates met the definition of synergy. Essential agreement of EUCAST and gradient concentration strip MICs at +/- 2 log

Topics: Agar; Antifungal Agents; Candida; Candidiasis; Colistin; Colony Count, Microbial; Drug Synergism; Humans; Microbial Sensitivity Tests; Nitriles; Pyridines; Triazoles

Candida species is implicated in a wide array of clinical infections. Speciation of Candida strains is of prime importance in the epidemiological survey and laboratory diagnosis as there is an upswing of antifungal resistance and changing trends in the antifungal resistance pattern among C. albicans and non albicans Candida. Varied phenotypic methods are available for the identification of Candida species which vary in principles and cost factors. Chromogenic agar medium (HiCrome Candida differential agar) is one of the preferred phenotypic methods in limited resource laboratories. Hence, this study was aimed to assess the reliability of HiCrome Candida differential agar, M1297A (HiMedia) in the identification of Candida species compared polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Oral Candida isolates (n = 194) were inoculated onto HiCrome Candida differential agar and the potential of Candida differential Agar was compared with PCR-RFLP.. The results were not in agreement with PCR-RFLP. Percentage of disagreement was 40.2, 50.0, 100.0 and 25.0 for Candida albicans, Candida krusei, Candida glabrata and Candida tropicalis respectively. PCR-RFLP demonstrated a very high discriminatory power in the identification of Candida species compared to agar.

Topics: Agar; Candida; Candida albicans; Candida glabrata; Candida tropicalis; Candidiasis; Chromogenic Compounds; Culture Media; Humans; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Reproducibility of Results; Sensitivity and Specificity; Species Specificity

The germ tube test (GTT) is inexpensive, easy, and well-defined test that differentiates Candida albicans (excluding Candida dubliniensis and Candida africana) from other species. The aim of this study was to evaluate various serums (i.e., human, rabbit, horse, and fetal bovine serum) used in the GTT and Mueller-Hinton agar (MHA).. Fifty species isolated from various clinical samples that were defined as C. albicans by both conventional and DNA sequence analysis methods were included in the study. One to two colonies of C. albicans were mixed into 0.5-1 ml of fetal bovine serum, horse serum, rabbit serum, and human serum. Serums and MHA were incubated at 37°C for GTT. They were removed from the incubator and evaluated after 30 min, 1 h, 2 h, and 3 h of incubation. The GTT was accepted to be positive only if germ tube was 1/2 the width and 3 times the length of the parent yeast cell and with no constriction at the point of origin.. When the use of serums and MHA for GTT was statistically evaluated, according to the positive scoring, the best results were obtained with MHA and with rabbit, horse, and fetal bovine serum, respectively. The best definition over time statistically was the third hour.. It is suggested that inexpensive MHA is a fast, appropriate, and reliable medium for the probable diagnosis of GTT and C. albicans; however, additional studies are still needed to define other Candida species.

Topics: Agar; Animals; Candida albicans; Candidiasis; Cattle; Culture Media; Horses; Humans; Molecular Typing; Multiplex Polymerase Chain Reaction; Mycological Typing Techniques; Rabbits; Sensitivity and Specificity; Serum; Species Specificity

Fluconazole is an alternative for candidemic patients who are not critically ill. Fluconazole is mainly fungistatic and does not completely inhibit visual Candida albicans growth. We studied the usefulness of fluconazole-containing Sabouraud dextrose agar plates for detecting susceptibility to fluconazole in C. albicans.. Adjusted inocula of 19 isolates were transferred directly onto fluconazole-containing Sabouraud dextrose plates (concentrations ranging from 0.125 mg/L to 128 mg/L). The fluconazole MIC in fresh isolates and after growth on the fluconazole-containing plate at 128 mg/L was recorded following the EUCAST EDef 7.2 guidelines. Then isolates were classified according to their degree of trailing production, based on microdilution procedure.. All isolates were able to grow on all fluconazole-containing plates, even those isolates susceptible to fluconazole. In fact, we selected isolates with different degrees of trailing based on microdilution procedures. 50% of isolates classified as heavy trailers, 35.71% as moderate trailers, and 14.28% as slight trailers.. The use of fluconazole-containing Sabouraud dextrose agar plates was not a reliable method to detect fluconazole susceptibility in C. albicans isolates; growth of the isolates was a trailing effect rather than true resistance.

Topics: Agar; Antifungal Agents; Candida albicans; Candidiasis; Culture Media; Fluconazole; Glucose; Microbial Sensitivity Tests; Reproducibility of Results

To evaluate the effects of two commercial silicone hydrogel contact lenses (CLs) soaked with natamycin (NA) or fluconazole (FL) on the growth of Candida albicans in an in vitro eye model.. Three-D printed molds were used as a cast for making eye-shaped models comprising potato dextrose agar. Senofilcon A (SA) and lotrafilcon B (LB) CLs were incubated with either 2 mL of NA or FL at a concentration of 1 mg/mL for 24 hr. To simulate a fungal infection, the eye models were coated with C. albicans. The drug-soaked lenses were placed on top of the eye models. Seven experimental conditions were examined: (1) NA-SA, (2) NA-LB, (3) FL-SA, (4) FL-LB, (5) SA, (6) LB, and (7) control-no lens. At specified time points (t=1, 8, 16, 24, 48 hr), the agar eyes from each experimental condition were removed from the incubator and photographed. The yeast cells from the 24 and 48 hr time point were also analyzed using light microscopy.. At 24 and 48 hr, there was considerable growth observed for all conditions except for the NA-SA and NA-LB conditions. When observed under the microscope at 24 and 48 hr, the morphology of the yeast cells in the FL-SA and SA condition were similar to that of the control (oval shaped). There was limited hyphae growth observed for LB and significant visible hyphae growth for the NA-LB group. For NA-SA, NA-LB, and FL-LB groups, the cells were significantly smaller compared with the control.. For NA-SA and NA-LB, there was limited growth of C. albicans observed on the eye models even after 48 hr. Under the microscope, the cell morphology differ noticeably between each testing condition, and is dependent on drug-lens combinations.

Topics: Agar; Antifungal Agents; Candida albicans; Candidiasis; Contact Lenses, Hydrophilic; Drug Delivery Systems; Eye Infections, Fungal; Fluconazole; Humans; Hydrogels; Keratitis; Models, Biological; Natamycin; Silicone Elastomers

Topics: Agar; Antifungal Agents; Azoles; Biological Assay; Candida; Candidiasis; Caspofungin; Combined Modality Therapy; Echinocandins; Humans; Lipopeptides; Microbial Sensitivity Tests

The aim of this study is to compare conventional methods, chromogenic agar, [corrected] VITEK2 YST card and VITEK®MS system for the identification of Candida strains isolated from blood cultures. Fifty-four strains were identified according to conventional methods, chromogenic agar, [corrected] VITEK2 YST card and VITEK®MS. Sequencing was used as the reference method. The 54 strains included 32 Candida parapsilosis, 19 Candida albicans, 1 Candida glabrata and 2 Candida tropicalis according to the reference method. One C. albicans and one C. glabrata isolate were misidentified as C. parapsilosis by chromogenic agar. [corrected]. Two C. parapsilosis and three C. albicans isolates were misidentified by VITEK2 YST card. Chromogenic agar, [corrected] VITEK2 YST card and VITEK®MS identified correctly 96.2%, 90.7% and 100% of all strains, respectively. We found that the chromogenic agar, [corrected] VITEK2 YST card and VITEK®MS system are easy, rapid and accurate alternative methods for the identification of yeast species in the clinical microbiology laboratory.

Topics: Agar; Blood Culture; Candida; Candida albicans; Candida glabrata; Candida tropicalis; Candidiasis; Culture Media; Humans; Mycological Typing Techniques; Predictive Value of Tests; Sensitivity and Specificity

Topics: Agar; Cacao; Candida; Candida albicans; Candidiasis; Culture Media

Candida albicans, a dimorphic fungus, undergoes hyphal development in response to many different environmental cues, including growth in contact with a semi-solid matrix. C. albicans forms hyphae that invade agar when cells are embedded in or grown on the surface of agar, and the integral membrane protein Dfi1p is required for this activity. In addition, Dfi1p is required for full activation of mitogen activated protein kinase Cek1p during growth on agar. In this study, we identified a putative calmodulin binding motif in the C-terminal tail of Dfi1p. This region of Dfi1p bound to calmodulin in vitro, and mutations that affected this region affected both calmodulin binding in vitro and invasive filamentation when incorporated into the full length Dfi1p protein. Moreover, increasing intracellular calcium levels led to calcium-dependent, Dfi1p-dependent Cek1p activation. We propose that conformational changes in Dfi1p in response to environmental conditions encountered during growth allow the protein to bind calmodulin and initiate a signaling cascade that activates Cek1p.

Topics: Agar; Amino Acid Motifs; Amino Acid Sequence; Animals; Antifungal Agents; Calcium; Calmodulin; Candida albicans; Candidiasis; Cell Wall; Disease Models, Animal; Fungal Proteins; Hyphae; Intracellular Space; Mice; Microbial Sensitivity Tests; Molecular Sequence Data; Mutant Proteins; Mutation; Protein Binding; Signal Transduction; Virulence

Although haemolytic factor is known to be a putative virulence factor contributing to pathogenicity in Candida species, its production by Candida tropicalis is poorly understood. In this study, we analysed the culture conditions under which C. tropicalis can display haemolytic factor on plate assay and the secretion of haemolytic factor in liquid medium by clinical isolates obtained from different specimens. All the tested isolates exhibited an internal translucent ring, resembling beta-haemolysis, surrounding by a peripheral greenish-grey halo on sheep blood agar medium. Similar haemolytic pattern was observed on human blood enriched medium. Furthermore, incubation either under normal atmosphere or under increased CO(2) had no effect on haemolysis. Overall, no differences were observed on beta-haemolytic activities (P > 0.05) among tested isolates of C. tropicalis. In glucose-limited medium (RPMI 1640 with 0.2% glucose), none of the isolates induced haemolysis on red blood cells. Similarly to found on plate assays, there were no significant differences (P > 0.05) in the activity of secreted haemolytic factor in liquid medium among C. tropicalis isolates. However, after growth, the number of yeast cells varied among isolates revealing different efficiencies of haemolytic factor production. Haemolytic activity was neither inhibited by heat treatment (100 °C) nor by the addition of pepstatin A. The obtained results extend our knowledge about haemolytic factor production by Candida species.

Topics: Agar; Animals; Candida tropicalis; Candidiasis; Carbon Dioxide; Culture Media; Erythrocytes; Hemolysin Proteins; Hot Temperature; Humans; Mycology; Sheep

The diagnosis of candida balanitis should be based upon both clinical and mycological data. The procedure of material collection is a critical issue to confirm or rule out the clinical diagnosis of candida balanitis.. To compare direct impression of the glans on the agar surface of solid culture media with the collection of genital exudates with cotton swab for the diagnosis of candida balanitis.. A prospective cross-sectional study was carried out during a 36-month period. Sexually transmitted disease clinic attendees with balanitis and asymptomatic men were included. Specimens for yeast culture were collected from the glans penis and inner preputial layer using the direct impression on CHROMagar candida medium and by swabbing with a sterile cotton swab.. Among 478 men enrolled, 189 had balanitis. The prevalence of candida balanitis was 17.8% (85/478) confirmed after culture by direct impression; the swab method detected only 54/85 (63.5%) of these men. Of the 289 asymptomatic men, 36 (12.5%) yielded Candida spp; the swab method detected only 38.9% of these men. The risk of having candida balanitis is 8.9 (IC 95% 2.48 to 32.04) whenever the number of candida colonies recovered by direct impression was greater than 10.. Direct impression on CHROMagar candida medium resulted in the highest Candida spp recovery rate. More than 10 colonies yielded by impression culture were statistically associated with candida balanitis. This method shows the ideal profile for sampling the male genital area for yeasts and should be included in the management of balanitis.

Topics: Adolescent; Adult; Agar; Aged; Aged, 80 and over; Balanitis; Candida; Candidiasis; Colony Count, Microbial; Cross-Sectional Studies; Culture Media; Exudates and Transudates; Humans; Male; Middle Aged; Penis; Specimen Handling; Young Adult

We examined the utility of agar dilution to screen yeasts for reduced susceptibility to several newer antifungal drugs including echinocandins and azoles. We compared agar dilution susceptibility screening with the Clinical and Laboratory Standards Institute (CLSI) method for Candida isolates. We added echinocandins and azoles to CHROMagar Candida medium prior to its solidification. Assessment of resistance was based on growth characteristics, wherein decreased colony size in the presence of antifungal drugs was used as an indicator of susceptibility. Clinical Candida isolates of C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. guilliermondii, C. lusitaniae, C. rugosa and C. dubliniensis were screened for drug susceptibility. Overall, antifungal susceptibility of the yeasts to anidulafungin, caspofungin, micafungin, posaconazole and voriconazole, determined using CHROMagar agar dilution, were shown to be 96, 80, 94, 90 and 97% as accurate, respectively, as those determined by the CLSI method, i.e., within one tube dilution of CLSI MICs. Categorical errors by percentage had a broader range. Major errors noted with anidulafungin, caspofungin and micafungin were 3, 6 and 0%, respectively, while very major errors were 15, 55 and 38%, respectively. Major errors with posaconazole and voriconazole were 12 and 0%, respectively, while very major errors were 0 and 22%, respectively, compared to CLSI standards. Most of the assessment errors were found with C. glabrata and C. parapsilosis. Agar dilution screening for drug susceptibility with the current panel of antifungal drugs is rapid, accurate and effective. However, the determination of resistance or non-susceptibility in yeasts may be more problematic, and may be species dependent.

Topics: Agar; Antifungal Agents; Candida; Candidiasis; Culture Media; Humans; Microbial Sensitivity Tests

Fungal infections caused by yeasts have increased during the last decades and invasive forms represent a serious problem for human health. Candida albicans is the species most frequently isolated from clinical samples. However, other emerging yeast pathogens are increasingly responsible for mycotic infections, and some of them are resistant to some antifungal drugs. Consequently, it is necessary to have methods that can provide a rapid presumptive identification at species level. Numerous chromogenic agar media have been shown to be of value as diagnostic tools. We have compared a chromogenic medium, Brilliance Candida Agar, with CHROMagar Candida, the chromogenic medium most used in our country. A multicentre study was conducted in 16 Hospitals belonging to the Mycology Net of Buenos Aires City Government. A total of 240 yeast isolates were included in this research. The new chromogenic agar showed results very similar to those obtained with CHROMagar Candida.

Topics: Agar; Argentina; Candida; Candidiasis; Chromogenic Compounds; Culture Media; Humans; Mycological Typing Techniques; Predictive Value of Tests; Sensitivity and Specificity; Species Specificity

Candida dubliniensis pathogenic species, which shares many phenotypic features with C. albicans, may be misidentified in the microbiology laboratory. The growth on DRBC agar at 25 degrees C was shown to be a new tool for differentiation between C. dubliniensis and C. albicans. All 27 isolates of C. dubliniensis showed in this medium rough colonies (peripheral hyphal fringes) and abundant chlamydospore production, while all 103 isolates of C. albicans showed smooth colonies without fringes or chlamydospores. DRBC agar allowed the differentiation of C. albicans from C. dubliniensis with 100 % sensitivity and specificity.

Topics: Agar; Candida; Candidiasis; Culture Media; Humans; Mycological Typing Techniques

A new method for quantification of yeast invasion into the agar medium was developed. Classical agar-invasion assays have been the methods of choice for determination of yeast invasion, but their main disadvantage is the lack of quantification. Our new Quantitative yeast agar-invasion test allows for quantitative measurements and enables sorting strains by their degree of invasiveness. The invasion abilities were measured for 10 clinical and non-clinical Saccharomyces cerevisiae strains and a strain of Candida albicans. Finally, the correlation between the degrees of strains invasiveness and their reported virulence was observed, proposing our assay as a method for quick determination of yeast virulence potential.

Topics: Agar; Candida albicans; Candidiasis; Colony Count, Microbial; Culture Media; Mycology; Saccharomyces cerevisiae; Statistics as Topic; Virulence; Yeasts

Caspofungin (CSP) susceptibilities of Candida albicans, as determined by broth microdilution methods, have not been found to be related to azole susceptibilities or resistance. In contrast, it has been observed that azole-resistant clinical isolates that overexpress the efflux pump gene CDR2 are less susceptible to CSP when tested using an agar dilution method commonly employed with Saccharomyces cerevisiae. The goal of this study was to further understand the effects of azole resistance mechanisms on CSP susceptibility testing. A collection of 69 isolates exhibiting known mechanisms of azole resistance and resistance-associated phenotypes were analyzed by broth microdilution methods to determine standard minimum inhibitory concentrations (MICs) for CSP. The same isolates were then analyzed as to their MIC to CSP by Etest strips, an agar-based method that has been shown generally to be comparable to broth methods. The MICs found with both methods were not significantly different. However, a collection of strains overexpressing the efflux pump CDR2 did exhibit a spectrum of CSP susceptibilities when examined by agar dilution susceptibility tests, ranging from standard to reduced susceptibilities. This work demonstrated that a change in CSP susceptibility with CDR2 overexpressing cells in agar dilution studies is a variable phenotype and it is not the result of growth conditions (i.e., broth versus agar).

Topics: Agar; Antifungal Agents; Azoles; Candida albicans; Candidiasis; Caspofungin; Drug Resistance, Fungal; Echinocandins; Fungal Proteins; Humans; Lipopeptides; Metallothionein; Microbial Sensitivity Tests

This study analyzed the correlation between the results obtained through two microdilution methods: Clinical and Laboratory Standard Institute (CLSI) (M27-A2) and European Committee on Antibiotic Susceptibility Testing (EUCAST) (document E. Dis. 7.1) and an agar base method Etest for determining minimmun inhibitory concentration (MIC) for amphotericin B and fluconazole against 30 clinical isolates of Candida spp. The agreement between Etest, CLSI and EUCAST MICs within +/- 2 log2 dilutions was higher for amphotericin B than for fluconazole However, Pearson correlation demonstrated a greater agreement for fluconazole. The categorical agreement between MICs provided by the Etest/ CLSI and Etest/EUCAST methodologies was high for both amphotericin B (100%) and fluconazole (> or = 96.66%). This study demonstrated the adequacy of Etest method using Mueller Hinton agar to evaluate amphotericin B and fluconazole susceptibility of clinical isolates of Candida spp.

Topics: Agar; Amphotericin B; Antifungal Agents; Candida; Candidiasis; Culture Media; Diffusion; Fluconazole; Microbial Sensitivity Tests

Candida dubliniensis is a recently described pathogenic species which shares many phenotypic features with Candida albicans and therefore, may be misidentified in microbiological laboratories. Because molecular methods can be onerous and unfeasible in routine mycological laboratories with restricted budgets such as those in developing countries, phenotypic techniques have been encouraged in the development of differential media for the presumptive identification of these species. We examined the colony morphology and chlamydospore production of 30 C. dubliniensis isolates and 100 C. albicans isolates on two new proposed media: rosemary (Rosmarinus officinalis) extract agar (REA) and oregano (Origanum vulgare) extract agar (OEA). These substrates are traditionally used as spices and medicinal herbs. In both of these media, all C. dubliniensis isolates (100%) showed rough colonies with peripheral hyphal fringes and abundant chlamydospores after 24 to 48 hr of incubation at 25 degrees C. In contrast, under the same conditions, all isolates of C. albicans (100%) showed smooth colonies without hyphal fringes or chlamydospores. In conclusion, REA and OEA offer a simple, rapid, and inexpensive screening media for the differentiation of C. albicans and C. dubliniensis.

Topics: Agar; Candida albicans; Candidiasis; Culture Media; Diagnosis, Differential; Female; Humans; Mycological Typing Techniques; Origanum; Plant Extracts; Prohibitins; Rosmarinus

We performed Etest, disk diffusion, and broth microdilution susceptibility testing of 2,171 clinical isolates of Candida spp. against posaconazole. By using provisional breakpoints for comparison purposes only, the categorical agreement between the agar-based methods and broth microdilution results ranged from 93 to 98%, with <1% very major errors. The essential agreement (within 2 well dilutions) between the Etest and broth microdilution methods was 94%. These agar-based methods hold promise as simple and reliable methods for determination of the posaconzole susceptibilities of Candida spp.

Topics: Agar; Antifungal Agents; Candida; Candidiasis; Culture Media; Disk Diffusion Antimicrobial Tests; Humans; Microbial Sensitivity Tests; Triazoles

Candida lusitaniae is an opportunistic yeast pathogen that has the ability to develop resistance to amphotericin B (AmB). The mechanism(s) for this resistance is not well understood, although there are data supporting mutations in sterol pathways and other data supporting phenotypic switching (PS). The goal of this study was to determine whether C. lusitaniae has a PS system and to characterize any phenotypes, including any changes in AmB MICs. When 10(4) CFU of an AmB-resistant (MIC of 16 to 32 microg/ml) clinical strain was plated on yeast-peptone-dextrose (YPD) agar with 1 mM CuSO(4), three colony colors were observed: light brown (LB) >> dark brown (DB) > white (W), similar to the result for Candida glabrata. Switching did occur with high AmB resistance (MIC of 256 microg/ml) being associated with W, whereas LB and DB colonies had MICs of 2 to 8 microg/ml and 2 to 16 microg/ml, respectively. Filamentation (pseudohyphae) was associated with DB colonies. All phenotypes occurred spontaneously with greater frequency ( approximately 10(-2) to 10(-4)) than spontaneous mutations, and all phenotypes were reversible, fulfilling the two PS criteria. High AmB MICs were always associated with W colonies but not with all W colonies. Detection of PS on YPD-CuSO(4) is also similar to that in Candida glabrata, and we hypothesize that this is due to similarities in metallothionein gene expression. Phenotypic switching represents a key strategy in C. lusitaniae that confers a selective advantage during environmental challenges, including the ability to switch to AmB resistance.

Topics: Agar; Amphotericin B; Candida; Candidiasis; Copper Sulfate; Culture Media; Drug Resistance, Fungal; Gene Expression Regulation, Fungal; Microbial Sensitivity Tests; Phenotype

Invasive diagnostic and therapeutic methods, widespread antibiotic therapy and rising percent of the immunocompromised patients cause incrementation of frequency of occurrence of the yeast infection. C. albicans is the most commonly isolated species of Candida from clinical samples. However, recently growth of frequency of isolation Candida non - albicans from clinical specimens have been observed. Yeast-like fungi different from C. albicans have become serious clinical problem. Conventional methods of identification of the yeast-like fungi carry away a lot time enough. Employment of chromogenic agar shortens latency on result. We decided to examine the usefulness ofAgar Candida ID2 (CAN2) (bioMérieux) in the identification of Candida species. The subjects within the study were 146 of Candida spp strains which were isolated from the clinical specimens of patients hospitalized at the University Hospital in Bydgoszcz. Germ tube test. Api 20C AUX test (bioMérieux) and Agar Candida ID2 (bioMérieux) were used. We have ascertained correspondence of identifying species amounted to 82.2% of analyzed Candida species between API 20C AUX test and kind of growth on CAN2 medium. Divergence of results received between CAN2 medium and API 20C AUX test suggests necessity of conducting of verification data with other methods. In conclusion, our study shows that Agar Candida ID2 is an effective medium for the isolation yeast-like fungi and in preliminary identification of Candida species direct from clinical materials.

Topics: Agar; Candida; Candidiasis; Cell Culture Techniques; Chromogenic Compounds; Culture Media; Female; Humans; Male; Mycological Typing Techniques; Reproducibility of Results; Sensitivity and Specificity; Species Specificity

In 1995, Candida dubliniensis was described as a new species in the genus Candida. Its close relationship to C. albicans has proved problematic in the identification of C. dubliniensis in clinical specimens. The objective of this study was to determine if reproducible differentiation between both species can be obtained by phenotypic assays. Therefore, 100 strains from 86 patients with the ability to produce chlamydospores were examined with different methods including API ID 32 C, colour development on CHROMagar, chlamydospore formation on Staib agar, growth at different temperatures and germ tube formation at 39 degrees C. Additionally, polymerase chain reaction (PCR) was used as gold standard. Six of the investigated strains were C. dubliniensis. The results suggest that there is still no single phenotypic method satisfactory to distinguish between C. albicans and C. dubliniensis.

Topics: Agar; Candida; Candida albicans; Candidiasis; Chromogenic Compounds; Culture Media; DNA, Fungal; Female; Humans; Male; Mycological Typing Techniques; Phenotype; Polymerase Chain Reaction; Spores, Fungal

An evaluation was performed on 95 blood cultures positive for Candida spp. to determine the correlation of direct susceptibility testing of fluconazole versus both standardized disk diffusion and MIC methods. For direct testing, an aliquot taken from BD BACTEC Plus and/or BD BACTEC Lytic/10 bottles (Becton Dickinson [BD], Sparks, MD) positive by gram stain for yeast was subcultured to CHROMagar Candida (BD), and a 25-microg fluconazole disk (BD) was placed on the plate. The area of growth inhibition surrounding the disk was measured at 24 and 48 h. In addition, a subculture of the isolate was tested by a microdilution MIC using YeastOne (TREK Diagnostics Systems Inc., OH) and disk diffusion (NCCLS M44-A) using a standardized inoculum plated onto CHROMagar Candida as well as Mueller-Hinton agar to which 2% glucose and 0.5 microg/ml methylene blue dye was added (MH-GMB). The categorical interpretation derived from the MIC was used as the reference to which the disk diffusion results were compared. There were a total of 41 Candida albicans, 23 Candida glabrata, 20 Candida parapsilosis, 9 Candida tropicalis, and 1 each of Candida krusei and Candida lusitaniae tested. At 24 h there was full agreement among the methods for all C. albicans, C. tropicalis, C. lusitaniae, and C. krusei isolates. For the C. parapsilosis isolates at 24 h there was one very major discrepancy using the direct CHROMagar and one major error with the standardized MH-GMB. The majority of the errors were seen at 24 h with the C. glabrata isolates. Of the 23 C. glabrata isolates at 24 h by direct CHROMagar, there were 10 minor and 1 very major error; by MH-GMB there were 12 minor and 2 very major errors; and by standardized CHROMagar Candida there were 13 minor and 2 major errors. There were no very major errors with C. glabrata when all plates were read at 48 h. At 24 h by the direct and standardized CHROMagar the majority of C. glabrata isolates were more resistant, whereas by MH-GMB they were more susceptible than the reference MIC interpretation. In summary, subculturing yeast directly from blood cultures onto CHROMagar to which a fluconazole disk has been added may provide a presumptive identification at 24 h and, with the exception of C. glabrata, was able to predict the susceptibility to fluconazole with the majority of Candida isolates examined in this evaluation.

Topics: Agar; Antifungal Agents; Blood; Candida; Candidiasis; Chromogenic Compounds; Culture Media; Fluconazole; Fungemia; Humans; Microbial Sensitivity Tests

This paper describes a simple and rapid method for the differentiation of Candida albicans from other yeast species in primary cultures based on colonial morphology following incubation in carbon dioxide. The technique has superior sensitivity to the traditional germ-tube method and requires no additional laboratory tests. In a busy laboratory, this can result in significant savings in cost and time, as well as improvements in patient care.

Topics: Agar; Candida albicans; Candidiasis; Carbon Dioxide; Colony Count, Microbial; Cost Savings; Culture Media; Diagnosis, Differential; DNA, Fungal; Hot Temperature; Humans; Mycological Typing Techniques; Phenotype; Sensitivity and Specificity; Time Factors

A sunflower (Helianthus annuus) seed husk agar medium has been developed and evaluated for differentiation of Candida dubliniensis from Candida albicans on the basis of colony morphology and chlamydospore production. All C. dubliniensis isolates (n=40) produced rough colonies with hyphal fringes and abundant chlamydospores whereas 101 of 105 (96.2%) C. albicans isolates produced smooth colonies with no evidence of chlamydospore production. Since this medium is free from oil droplets, chlamydospores can be examined with greater clarity by Dalmau plate technique. This medium provides a simple and cost-effective tool for the presumptive differentiation of C. dubliniensis from C. albicans and is particularly suited for clinical microbiology laboratories where biochemical or molecular methods for the differentiation of these two species are not available.

Topics: Agar; Candida albicans; Candidiasis; Diagnosis, Differential; Helianthus; Humans; Microbiological Techniques; Seeds

CHROMagar Candida medium is used for the isolation and identification of Candida species, but it does not differentiate Candida albicans from Candida dubliniensis. This differentiation can be achieved by using Pal's agar, which cannot be used in primary isolation. We have combined both media to obtain a new medium that can be used for the isolation and identification of C. dubliniensis in primary cultures.

Topics: Agar; Candida; Candidiasis; Culture Media; Humans; Mycological Typing Techniques; Species Specificity

Candida dubliniensis is an emerging yeast pathogen isolated mainly from immunocompromised patients. As molecular tests are currently unsuitable for use in routine diagnostic laboratories, we compared a variety of phenotypic techniques for differentiating C. albicans and C. dubliniensis. The tests included: colony colour on CHROMagar Candida medium; growth at 37 degrees C and 45 degrees C; ability to produce germ tubes and chlamydospores; and the Auxacolor system. The organisms included 105 isolates previously identified as C. albicans, 10 reference strains of C. albicans, 2 reference strains of C. dubliniensis and 102 fresh clinical isolates identified as C. albicans. None of the tests alone was satisfactory but a combination of 3 tests may be suitable for presumptive identification of C. dubliniensis.

Topics: Agar; AIDS-Related Opportunistic Infections; Candida; Candida albicans; Candidiasis; Chromogenic Compounds; Color; Communicable Diseases, Emerging; Culture Media; Culture Techniques; Diagnosis, Differential; DNA, Fungal; Drug Resistance, Fungal; Humans; Immunocompromised Host; Mycological Typing Techniques; Phenotype; Sensitivity and Specificity; Temperature; Time Factors

This study evaluated sunflower (Helianthus annuus) seed agar (SSA) for differentiation of Candida dubliniensis from Candida albicans on the basis of colony characteristics and chlamydospore production. Simplified SSA without creatinine and KH(2)PO(4) was also used. On both media, C. dubliniensis isolates (n = 25) developed rough colonies and formed abundant chlamydospores after incubation for 24-48 h at 28 degrees C, while C. albicans isolates (n = 53) showed smooth colonies with no evidence of chlamydospore formation. Cryptococcus neoformans isolates (n = 10) formed brown colonies on both media. Simplified SSA offers a simple and inexpensive tool for presumptive differentiation of C. dubliniensis from C. albicans in clinical microbiology laboratories.

Topics: Agar; Candida; Candida albicans; Candidiasis; Culture Media; Helianthus; Humans; Mycological Typing Techniques; Seeds; Spores, Fungal

Isolates of Candida dubliniensis may be misidentified as Candida albicans in microbiological laboratories if only the germ tube and/or the chlamydospore test is used for identification to the species level. In this study, we have evaluated the efficacy of tobacco agar for the differentiation of C. dubliniensis from C. albicans. On this medium at 28 degrees C, all 30 C. dubliniensis isolates produced yellowish-brown colonies with hyphal fringes and abundant chlamydospores, whereas 54 C. albicans isolates formed smooth, white-to-cream-colored colonies with no chlamydospore production. This medium provides a simple tool for presumptive differentiation of C. dubliniensis from C. albicans.

Topics: Agar; Candida; Candida albicans; Candidiasis; Culture Media; Humans; Mycological Typing Techniques; Nicotiana; Species Specificity

The performance of the Etest using Mueller-Hinton agar supplemented with glucose (2%) and methylene blue (0.5 microg/ml) (MH-GMB) for amphotericin B susceptibility testing of 4,936 isolates of Candida spp. was assessed against that of Etest using RPMI agar with 2% glucose (RPG). MICs were determined by Etest in both media for all 4,936 isolates and were read after incubation for 48 h at 35 degrees C. The Candida isolates included C. albicans (n = 2,728), C. glabrata (n = 722), C. parapsilosis (n = 666), C. tropicalis (n = 528), C. krusei (n = 143), C. lusitaniae (n = 54), C. guilliermondii (n = 39), C. pelliculosa (n = 17), C. kefyr (n = 15), C. rugosa (n = 11), C. dubliniensis (n = 5), C. zeylanoides (n = 4), C. lipolytica (n = 3), and C. famata (n = 1). The Etest results with MH-GMB correlated well with those with RPG. Overall agreement was 92.9%, and agreements for individual species were as follows: C. lusitaniae, 98.1%; C. albicans, 95.1%; C. glabrata, 94.3%; C. krusei, 91.6%; C. parapsilosis, 86.6%; and C. tropicalis, 86.4%. The Etest method using MH-GMB appears to be a useful method for determining amphotericin B susceptibilities of Candida species.

Topics: Agar; Amphotericin B; Antifungal Agents; Candida; Candidiasis; Culture Media; Glucose; Humans; Methylene Blue; Microbial Sensitivity Tests

The performances of the Etest and the disk diffusion methods for testing of the susceptibilities of 235 Candida glabrata isolates to fluconazole and voriconazole were compared with that of the National Committee for Clinical Laboratory Standards (NCCLS) approved standard broth microdilution (BMD) method. The NCCLS method used RPMI 1640 broth medium, and MICs were read after incubation for 48 h at 35 degrees C. Etest MICs were determined with RPMI 1640 agar containing 2% glucose (RPG agar) and with Mueller-Hinton agar containing 2% glucose and 0.5 microg of methylene blue per ml (MBE agar) and were read after incubation for 48 h at 35 degrees C. Disk diffusion testing was performed with MBE agar, 25-microg fluconazole disks, and 1- microg voriconazole disks and by incubation at 35 degrees C for 24 h. Overall agreements between the Etest and the BMD MICs obtained with RPG and MBE agars were 91 and 96%, respectively, for fluconazole and 93 and 95%, respectively, for voriconazole. Categorical agreements between the agar-based methods and BMD were 52.3 to 64.7% with fluconazole and 94.8 to 97.4% with voriconazole. The vast majority of the discrepancies by the disk diffusion and Etest methods with fluconazole were minor errors. The agar-based methods performed well in identifying isolates with resistance to fluconazole and decreased susceptibility to voriconazole.

Topics: Agar; Antifungal Agents; Candida glabrata; Candidiasis; Culture Media; Fluconazole; Fungemia; Humans; Microbial Sensitivity Tests; Pyrimidines; Quality Control; Triazoles; Voriconazole

CHROMagar Candida is a selective and differential chromogenic medium that has been shown to be useful for identification of Candida albicans, Candida krusei, Candida tropicalis, and perhaps Candida glabrata. Colony morphology and color have been well defined when CHROMagar Candida has been used to isolate yeast directly from clinical specimens, including stool, urine, respiratory, vaginal, oropharyngeal, and esophageal sources. Direct isolation of yeast on CHROMagar Candida from blood cultures has not been evaluated. We evaluated whether the color and colony characteristics produced by Candida spp. on CHROMagar Candida were altered when yeasts were isolated directly from blood cultures. Fifty clinical isolates of Candida were inoculated into aerobic and anaerobic blood culture bottles and incubated at 35 degrees C in an automated blood culture system. When growth was detected, an aliquot was removed and plated onto CHROMagar Candida. As a control, CHROMagar Candida plates were inoculated with the same isolate of yeast grown on Sabouraud dextrose agar simultaneously. No significant difference was detected in color or colony morphology between the blood and control isolates in any of the tested organisms. All C. albicans (n = 12), C. tropicalis (n = 12), C. glabrata (n = 9), and C. krusei (n = 5) isolates exhibited the expected species-specific colony characteristics and color, whether isolated directly from blood or from control cultures. CHROMagar Candida can be reliably used for direct isolation of yeast from blood cultures. Direct isolation could allow mycology laboratories to more rapidly identify Candida spp., enable clinicians to more quickly make antifungal agent selections, and potentially decrease patient morbidity and mortality.

Topics: Aerobiosis; Agar; Anaerobiosis; Blood; Candida; Candidiasis; Chromogenic Compounds; Culture Media; Fungemia; Humans; Microbiological Techniques

This study was undertaken to evaluate the utility of CHROMagar Candida medium for the identification of chlamydospore-negative Candida albicans. A total of 60 isolates including 45 chlamydospore-negative C. albicans, 10 chlamydospore-positive C. albicans (positive controls) and five non-C. albicans (negative controls) were investigated. On the basis of germ tube test, assimilation of trehalose (Tre), glucosamine (GlcN), N-acetyl glucosamine (GlcNAc), secretory aspartyl production and serotyping, the 45 chlamydospore-negative C. albicans isolates were assigned to the reported three groups. Eighteen isolates showing positive germ tube test, negative for the assimilation of Tre, GlcN/GlcNAc, strong producers of proteinase (2+) and assigned to serotype B belonged to group I. Whereas, the isolates in group II and group III showed common characteristics including assimilation of Tre, GlcN/GlcNAc, moderate production of proteinase (1+) and were serotype A, except for the fact that group II isolates were germ tube positive and group III isolates were negative. Using CHROMagar Candida medium, all the 45 chlamydospore-negative and 10 positive control isolates were accurately identified on the basis of characteristic green color at 37 degrees C for 48 h of incubation. On the other hand at an optimum incubation temperature of 37 degrees C none of the non-C. albicans (negative controls) showed characteristic green color thus yielding a 100% sensitivity and specificity. Isolates in group-I showed a slow growth rate and no visible growth was observed at 24 h, whereas, groups II, III and the control isolates showed visible growth at 24 h. Besides differences in growth rates, these isolates also varied in their characteristic colony color which gradually changed over a period of time. The results of this study clearly suggest that CHROMagar Candida medium is not only a simple, reliable and cost effective method for the identification of chlamydospore-negative atypical C. albicans, but can also be used to differentiate various groups of chlamydospore-negative C. albicans.

Topics: Adult; Agar; Candida albicans; Candidiasis; Chromogenic Compounds; Culture Media; Female; Humans; Mycological Typing Techniques; Spores, Fungal

Production of a hyphal fringe around colonies grown on Pal's agar (sunflower seed agar) at 30 degrees C for 48 to 72 h provides a simple means of discriminating between isolates of C. dubliniensis and C. albicans with 100% accuracy. Of 128 C. dubliniensis isolates tested on this medium, all produced a hyphal fringe. In contrast, none of the 124 C. albicans isolates tested produced a hyphal fringe. Pal's medium has the added advantage of being prepared from inexpensive, readily available seeds.

Topics: Agar; Candida; Candida albicans; Candidiasis; Culture Media; Humans; Mycological Typing Techniques

Albicans ID2 (bioMérieux, France) is a commercially available chromogenic medium that allows rapid and specific macroscopic identification of Candida albicans and facilitates the differentiation of species in mixed cultures. We compared it with the standard method for the identification of yeast species, the germ tube test (GT). This study involved 423 clinical isolates, including 163 C. albicans and 260 non-albicans yeasts. Sensitivity of Albicans ID2 agar plates regarding the identification of C. albicans were 98.2% after 48 h of incubation and specificity of 96.6%. This method using rapid enzymatic method shows the same similar sensitivity than the GT test The false negative rate (1.8%) for the GT test is consistent with that previously reported. None tests discriminated between C. albicans and C. dubliniensis isolates.

Topics: Agar; Candida albicans; Candidiasis; Cell Culture Techniques; Culture Media; Humans; Mycological Typing Techniques; Sensitivity and Specificity

The methods currently available for the identification of the pathogenic yeast Candida dubliniensis all have disadvantages in that they are time-consuming, expensive, and/or, in some cases, unreliable. In a recent study (P. Staib and J. Morschhäuser, Mycoses 42:521-524; 1999) of 14 C. dubliniensis and 11 C. albicans isolates, it was suggested that the ability of C. dubliniensis to produce rough colonies and chlamydospores (chlamydoconidia) on Staib agar (SA) provided a simple means of differentiating it from its close relative C. albicans. In the present investigation, we examined the colony morphology and chlamydospore production of 130 C. dubliniensis and 166 C. albicans isolates on SA and on the related defined medium caffeic acid-ferric citrate agar (CAF). All of the C. dubliniensis and C. albicans isolates produced chlamydospores on the control medium, i.e., rice-agar-Tween agar. However, while none of the C. albicans isolates produced chlamydospores on either SA or CAF, 85.4 and 83.8% of the C. dubliniensis isolates produced chlamydospores on SA and CAF, respectively. All of the C. albicans isolates grew as smooth, shiny colonies on SA after 48 to 72 h of incubation at 30 degrees C, while 97.7% of the C. dubliniensis isolates grew as rough colonies, many (65%) with a hyphal fringe. In contrast, 87.4% of the C. albicans and 93.8% of the C. dubliniensis isolates yielded rough colonies on CAF. Although the results of this study confirm that SA is a good medium for distinguishing between C. dubliniensis and C. albicans, we believe that discrimination between these two species is best achieved on the basis of colony morphology rather than chlamydospore production.

Topics: Agar; Caffeic Acids; Candida; Candida albicans; Candidiasis; Culture Media; Ferric Compounds; Humans; Spores, Fungal

All though extremely rare 10 years ago, antifungal drug resistance is becoming a major problem in certain populations, especially in those infected with HIV. This study was undertaken to study the resistance of Candida species isolated in our hospital to Fluconazole using Chrom agar Candida. The Candida strains which were routinely isolated from clinical specimens like blood, urine, sputum, pus, fluid and homograft isolates were included in the study. 142 Candida isolates were tested by using Chrom agar Candida incorporated with Fluconazole. 16 strains were found to be resistant to Fluconazole and 126 strains sensitive to Fluconazole. Nine were C. tropicalis, 3 C. krusei, 2 C. guillermondii, 1 Geotrichum candidum and one was an unidentified strain of Candida. The MIC of the 16 strains were done using RPMI 1640 medium by macro broth dilution method. MIC of 9 strains was 64 & > 64 ug/ml of 6 strains 32 ug/ml and 1 strain 16 ug/ml.

Topics: Agar; Antifungal Agents; Candida; Candidiasis; Drug Resistance, Fungal; Fluconazole; Humans; Microbial Sensitivity Tests

In many developing countries sheep and horse blood, the recommended blood supplements in bacteriological media, are not readily available, whereas pig and goat blood are. Therefore, this study examined the use of pig and goat blood as potential substitutes for sheep blood in blood-supplemented bacteriologic media commonly used in clinical microbiology laboratories. In general, the growth characteristics and colony morphologies of a wide range of aerobic and anaerobic bacteria and Candida albicans were similar on media containing pig, goat, and sheep blood, although differences were found. Enterococcus sp. uniformly produced alpha-hemolysis when incubated in CO(2), but in anaerobic conditions the hemolysis varied. In contrast, beta-hemolytic streptococci produced identical hemolytic reactions on all three media. Synergistic hemolysis was not observed on pig blood agar in the CAMP test nor on goat blood agar in the reverse CAMP test. The preparation of chocolate agar (heated) with pig blood required heating to a higher temperature than with sheep or goat blood to yield suitable growth of Haemophilus species. In general, we conclude that pig and goat blood are suitable alternatives to sheep blood for use in bacteriological media in settings where sheep and horse blood are not readily available.

Topics: Agar; Animals; Bacteria; Bacterial Infections; Bacteriological Techniques; Blood; Candida; Candidiasis; Culture Media; Goats; Humans; Sheep; Swine

To evaluate the activity of benzydamine, lidocaine, and bupivacaine, three drugs with local anesthetic activity, against Candida albicans and non-albicans strains and to clarify their mechanism of activity.. The minimal inhibitory concentration (MIC) was determined for 20 Candida strains (18 clinical isolates and two American Type Culture Collection strains). The fungistatic activity was studied with the fluorescent probe FUN-1 and observation under epifluorescence microscopy and flow cytometry. The fungicidal activity of the three drugs was assayed by viability counts. Membrane alterations induced in the yeast cells were evaluated by staining with propidium iodide, by quantitation of intracellular K+ leakage and by transmission electron microscopy of intact yeast cells and prepared spheroplasts.. The MIC ranged from 12.5-50.0 microg/mL, 5.0-40.0 mg/mL, and 2.5-10.0 mg/mL for benzydamine, lidocaine, and bupivacaine, respectively. The inhibitory activity of these concentrations could be detected with the fluorescent probe FUN-1 after incubation for 60 minutes. A very fast fungicidal activity was shown by 0.2, 50, and 30 mg/mL of benzydamine, lidocaine, and bupivacaine, respectively.. At lower concentrations, the tested drugs have a fungistatic activity, due to yeast metabolic impairment, while at higher concentrations they are fungicidal, due to direct damage to the cytoplasmic membrane.

Topics: Agar; Anesthetics, Local; Anti-Inflammatory Agents, Non-Steroidal; Antifungal Agents; Benzydamine; Bupivacaine; Candida; Candidiasis; Cell Membrane Permeability; Colony Count, Microbial; Female; Flow Cytometry; Humans; Lidocaine; Microbial Sensitivity Tests; Microscopy, Electron; Microscopy, Fluorescence; Time Factors

Performance of the Etest for voriconazole susceptibility testing of 312 isolates of Candida spp. was assessed against that of the National Committee for Clinical Laboratory Standards (NCCLS) microdilution broth method. The NCCLS method employed RPMI 1640 broth medium, and MICs were read after incubation for 48 h at 35 degrees C. Etest MICs were determined with RPMI agar containing 2% glucose (RPG), Casitone agar (CAS), and antibiotic medium 3 (AM3) agar and were read after incubation for 48 h at 35 degrees C. The Candida spp. isolates included C. albicans (n = 174), C. glabrata (n = 55), C. tropicalis (n = 31), C. parapsilosis (n = 39), C. krusei (n = 5), C. lusitaniae (n = 2), and C. guilliermondii (n = 6). The Etest results obtained using RPG correlated well with the reference MICs. Overall agreement ranged from 91% for C. glabrata to 100% for C. tropicalis, C. parapsilosis, C. guilliermondii, C. krusei, and C. lusitaniae. When CAS was used, agreement ranged from 80% for C. krusei to 100% for C. parapsilosis, C. guilliermondii, and C. lusitaniae. With AM3, agreement ranged from 58% for C. glabrata to 100% for C. lusitaniae and C. guilliermondii. The Etest method using RPG appears to be a useful method for determining voriconazole susceptibilities of Candida species.

Topics: Agar; Antifungal Agents; Candida; Candida albicans; Candidiasis; Culture Media; Humans; Microbial Sensitivity Tests; Pyrimidines; Triazoles; Voriconazole

The performance of the Etest for fluconazole susceptibility testing of 402 yeast isolates was assessed against the National Committee for Clinical Laboratory Standards (NCCLS) microdilution broth method. The NCCLS method employed RPMI 1640 broth medium, and MICs were read after incubation for 48 h at 35 degrees C. Etest MICs were determined with RPMI agar containing 2% glucose (RPG), Casitone agar (CAS), and Mueller-Hinton agar (MHA) and were read after incubation for 48 h at 35 degrees C. The yeast isolates included Candida albicans (n = 161), Candida glabrata (n = 41), Candida tropicalis (n = 35), Candida parapsilosis (n = 29), Candida krusei (n = 32), Candida lusitaniae (n = 31), Candida species (n = 19), Cryptococcus neoformans (n = 40), and miscellaneous yeast species (n = 14). The Etest results correlated well with reference MICs. Overall agreement was 94% with RPG, 97% with CAS, and 53% with MHA. When RPG was used, agreement ranged from 89% for Candida spp. to 100% for C. krusei. When CAS was utilized, agreement ranged from 93% for Cryptococcus neoformans to 100% for C. tropicalis, C. parapsilosis, C. lusitaniae, Candida spp., and miscellaneous yeast species. With MHA, agreement ranged from 17% for C. parapsilosis to 90% for C. krusei. Both RPG and CAS supported growth of all yeast species, whereas growth on MHA was comparatively weaker. Etest results were somewhat easier to read on CAS. The Etest method using either RPG or CAS, but not MHA, appears to be a viable alternative to the NCCLS reference method for determining fluconazole susceptibilities of yeasts.

Topics: Agar; Antifungal Agents; Candida; Candida albicans; Candidiasis; Culture Media; Fluconazole; Humans; Microbial Sensitivity Tests

Topics: Agar; Aminopeptidases; Candida albicans; Candidiasis; Chromogenic Compounds; Culture Media; Hexosaminidases; Humans; Mycology

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

A simple screening method for fluconazole susceptibility using CHROMagar Candida with fluconazole was compared with the National Committee for Clinical Laboratory Standards (NCCLS) macrobroth method. In this agar dilution method, susceptible Candida albicans colonies are smaller on medium with fluconazole than on fluconazole-free medium. Yeasts with decreased susceptibility have normal-sized colonies on medium containing fluconazole. On agar with 16 micrograms of fluconazole per ml, 32 of 34 strains with NCCLS MICs of > or = 16 micrograms/ml were correctly predicted, as were 66 of 68 with MICs of < 16, an agreement of 96%. On agar with 8 micrograms of fluconazole per ml, 38 of 41 isolates with MICs of > or = 8 were correctly predicted, as were 59 of 61 isolates with MICs of < 8, an agreement of 95%. This agar dilution methods appears to highly correlate with NCCLS macrobroth methods for detection of C. albicans and may be an effective screen for fluconazole susceptibility.

Topics: Agar; Antifungal Agents; Candida albicans; Candidiasis; Chromogenic Compounds; Evaluation Studies as Topic; Fluconazole; Humans; Microbial Sensitivity Tests; Reference Standards

The aim of the study was to evaluate the chromogenic agar plate CPS ID2 (bioMérieux) and determine its cost-benefit ratio.. A total of 2,193 urinary sediments were processed. The urine culture was carried out in CPS ID2 agar and in cystine-lactose electrolyte deficient (CLED) agar, when needed. Identification of the microorganisms was performed following standard microbiologic procedures through biochemical tests prepared in our laboratory. The identification, from CPS ID2 agar, by direct detection in medium of four metabolic activities: beta-glucuronidase, beta-glucosidase, deaminase, and indol production, was performed following to manufacturer's instructions.. A total of 289 urine cultures were positive, 18 were negative and 34 were contaminated samples. The identification, directly performed from the colonies detected in CPS ID2 agar, was correct in 96% of 166 Escherichia coli, in 92% of 24 Proteus mirabilis and in 97% of 38 enterococci. CPS ID2 agar exhibited 94% and 100% sensitivity and specificity, respectively in E. coli identification, 92% and 100% in P. mirabilis and 97% and 99% in Enterococcus. The use of this new media, CPS ID2, in our laboratory, implies a budgetary increment. However, if commercial galleries are used for routine identification, the cost will be reduced using this new media.. The CPS ID2 agar allows the isolation and direct identification of the most frequent urinary tract pathogens: E. coli, P. mirabilis and Enterococcus in primary isolation medium. Using this medium, bacteriologists will be able to save time and reagents when identifying the most common uropathogens. Furthermore, the use of this medium would reduce costs in some laboratories.

Topics: Agar; Amino Acid Oxidoreductases; Bacterial Proteins; Bacteriological Techniques; beta-Glucosidase; Candida albicans; Candidiasis; Chromogenic Compounds; Cost-Benefit Analysis; Culture Media; Enterobacteriaceae; Enterobacteriaceae Infections; Evaluation Studies as Topic; Glucuronidase; Gram-Negative Bacteria; Gram-Negative Bacterial Infections; Gram-Positive Bacteria; Gram-Positive Bacterial Infections; Humans; Indoles; L-Amino Acid Oxidase; Urinary Tract Infections; Urine

CHROMagar Candida is a novel, differential culture medium that is claimed to facilitate the isolation and presumptive identification of some clinically important yeast species. We evaluated the use of this medium with 726 yeast isolates, including 82 isolated directly on the medium from clinical material. After 2 days of incubation at 37 degrees C, 285 C. albicans isolates gave distinctive green colonies that were not seen with any of 441 other yeast isolates representing 21 different species. A total of 54 C. tropicalis isolates also developed distinctive dark blue-gray colonies with a halo of dark brownish purple in the surrounding agar. C. krusei isolates (n = 43) also formed highly characteristic rough, spreading colonies with pale pink centers and a white edge that was otherwise encountered only rarely with isolates of C. norvegensis. Trichosporon spp. (n = 34) formed small, pale colonies that became larger and characteristically rough with prolonged incubation. Most of the other 310 yeasts studied formed colonies with a color that ranged from white to pink to purple with a brownish tint. The only exceptions were found among isolates identified as Geotrichum sp. or Pichia sp., some of which formed colonies with a gray to blue color and which in two instances formed a green pigment or a dark halo in the agar. The specificity and sensitivity of the new medium for the presumptive identification of C. albicans, C. krusei, and C. tropicalis exceeded 99% for all three species. A blinded reading test involving four personnel and 57 yeast isolates representing nine clinically important species confirmed that colonial appearance after 48 h of incubation on CHROMagar Candida afforded the correct presumptive recognition of C. albicans, C. tropicalis, C, krusei, and Trichosporon spp. None of nine bacterial isolates grew on CHROMagar Candida within 72 h, and bacteria (Escherichia coli) grew from only 4 of 104 vaginal, 100 oral, and 99 anorectal swabs. The new medium supported the growth of 19 of 23 dermatophyte fungi tested and 41 of 43 other molds representing a broad range of fungal pathogens and contaminants. In parallel cultures of 348 clinical specimens set up on Sabourand agar and CHROMagar Candida, both media grew yeasts in the same 78 instances. CHROMagar Candida is recommended as a useful isolation medium capable of the presumptive identification of the yeast species most commonly isolated from clinical material and facilitating recognition of mixed yeas

Topics: Agar; Candida; Candida albicans; Candidiasis; Chromogenic Compounds; Culture Media; Evaluation Studies as Topic; Humans; Morphogenesis; Single-Blind Method

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

All Candida albicans isolates in Norwegian microbiological laboratories in 1991 judged clinically important (except vaginal isolates) were collected. The isolates were tested for susceptibility to fluconazole with an agar dilution test and a commercially available agar diffusion test. A total of 212 strains (95%) were susceptible to fluconazole, and MICs for most of the strains (92%) were < or = 1.56 micrograms/ml. The agar diffusion test using a 15-micrograms tablet and a 48-h incubation period separated resistant from susceptible strains with a wide margin. The only exception was a strain for which the MIC was 6.25 micrograms/ml. The difference in zone size between the resistant and the susceptible populations of strains was 11 mm. Accordingly, it appears that the agar diffusion test is an appropriate method for detecting fluconazole resistance. The 12 fluconazole-resistant isolates originated from eight AIDS patients with oral or esophageal Candida infections. Seven of the patients had been given fluconazole for 1 month or more, often as self medication. Four had infections that were clinically resistant to fluconazole; one additional patient responded only when the dose was increased. All isolates recovered from these patients were analyzed by multilocus enzyme electrophoresis. The 12 C. albicans isolates belonged to five electrophoretic types, but three of four patients attending one hospital had isolates belonging to one electrophoretic type. One possible explanation for this finding could be that a nosocomial spread of resistant strains has occurred.

Topics: Acquired Immunodeficiency Syndrome; Agar; Candida albicans; Candidiasis; Culture Media; Drug Resistance, Microbial; Fluconazole; Genotype; Humans; Microbial Sensitivity Tests; Norway

The trial of a newly developed dried selective medium for the isolation of fungi of the genus Candida, involving the inoculation of 103 museum yeast strains and 542 specimens of pathological material, has been carried out. The data obtained in this trial indicate that the proposed medium has advantages over wort agar and Sabouraud medium with antibiotics in the germination index, in the ability to ensure fungal growth after the inoculation of pathological material, and in selective properties. The medium ensures the intactness of the morpho-cultural, biochemical and serological properties of the fungi. The results of the trial recommend the newly developed preparation for laboratory practice for the isolation of yeast-like fungi from clinical material in the diagnosis of candidiasis, intestinal microflora disturbances, as well as for isolation of the fungi from various environmental objects.

Topics: Agar; Body Fluids; Candida; Candidiasis; Culture Media; Escherichia coli; Evaluation Studies as Topic; Humans; Proteus vulgaris; Staphylococcus aureus

A simple (2% oxgall) agar medium is described for the production of both germ tubes and chlamydospores by over 500 clinical isolates of Candida albicans. In comparison studies between the 2% oxgall agar and the more complex "cream of rice" infusion-oxgall-Tween 80 agar (RIOT), germ tubes were formed by 478 isolates in both media, by 9 isolates in only the oxgall medium and by 11 isolates in only the RIOT medium. Chlamydospores were formed by 481 isolates in both media, by 2 isolates in only the oxgall medium and by 9 isolates in only the RIOT medium. The data also show that both the germ tubes and chlamydospores were needed for the presumptive clinical identification of C. albicans.

Topics: Agar; Candida albicans; Candidiasis; Humans; Spores, Fungal

Topics: Agar; Antibodies, Fungal; Antigens, Fungal; Candida albicans; Candidiasis; Gels; Humans; Polyethylene Glycols; Precipitin Tests

Topics: Adolescent; Agar; Aged; Amphotericin B; Candida albicans; Candidiasis; Cystitis; Female; Humans; Male; Middle Aged; Urine

Topics: Adult; Agar; Candidiasis; Cornea; Eye Injuries; Glucose; Humans; Male; Ontario; Pseudallescheria; Spores, Fungal

Topics: Agar; Agglutination Tests; Animals; Antibodies; Candidiasis; Cryptococcosis; Culture Media; Culture Techniques; Dog Diseases; Dogs; Germany, West; Horse Diseases; Horses; Methods

Topics: Administration, Oral; Adolescent; Adult; Agar; Aged; Candidiasis; Cephalosporins; Escherichia coli; Escherichia coli Infections; Female; Haemophilus Infections; Haemophilus influenzae; Humans; Klebsiella; Klebsiella Infections; Male; Microbial Sensitivity Tests; Middle Aged; Nausea; Proteus; Proteus Infections; Staphylococcal Infections; Streptococcal Infections; Streptococcus; Streptococcus pneumoniae; Streptococcus pyogenes; Urinary Tract Infections; Vulvovaginitis

Topics: Agar; Agglutination Tests; Animals; Brain; Candida; Candidiasis; Female; Gels; Haplorhini; Immune Sera; Immunity; Immunization Schedule; Immunization, Passive; Injections, Intramuscular; Injections, Intravenous; Kidney; Male; Myocardium; Precipitin Tests; Rabbits

Topics: Agar; Anti-Bacterial Agents; Candida; Candidiasis; Child; Humans; Immunodiffusion; Male; Middle Aged; Nystatin; Precipitin Tests

Topics: Agar; Amphotericin B; Candida; Candidiasis; Endocarditis; Fluorescent Antibody Technique; France; Humans; Immunodiffusion; Immunoelectrophoresis; Nystatin; Precipitins; Sepsis

Topics: Agar; Amebicides; Antifungal Agents; Aspergillosis; Candidiasis; Dermatomycoses; Drug Synergism; Humans; Microsporum; Quinolines; Resorcinols; Tinea

Nero, L. C. (Illinois Institute of Technology, Chicago), Mae G. Tarver, and L. R. Hedrick. Growth of Acanthamoeba castellani with the yeast Torulopsis famata. J. Bacteriol. 87:220-225. 1964.-Acanthamoeba castellani and Torulopsis famata cells were cultured together for 10 days at three different temperatures: 18, 25, and 32 C. In the 18- and 25-C series, the yeast population reached a peak within 3 to 5 days and then declined for the next 5 days. This decrease in yeast population corresponded with an increase in the development of amoebae, which first appeared as vegetative cells and later transformed themselves into cysts. The changes in population were determined by counts in a hemocytometer. The vegetative amoeba population seldom exceeded 4 x 10(7) cells per bottle culture, but the encysted population approached 16 x 10(7) cysts per bottle culture within 10 days of incubation. The amoebae actively ingested the yeasts and used these cells as their principal source of energy. At 18 C each amoeba consumed 70 yeast cells per day; at 25 C each amoeba ingested 35 yeast cells per day. In the 32-C series, both the yeast and the amoebae had slow growth rates. The development of cysts corresponded with the growth of the yeast population without the rapid increase in yeast population prior to the growth of the amoebic cells.

Topics: Acanthamoeba castellanii; Agar; Amoeba; Animals; Candida; Candida glabrata; Candidiasis; Cryptococcus; Invertebrates; Research; Saccharomyces cerevisiae; Yeasts