Search

A 13-month prospective multicenter study including 44 hospitals was carried out to evaluate the epidemiology of Candida parapsilosis complex candidemia in Spain. Susceptibility to amphotericin B, flucytosine, fluconazole, itraconazole, voriconazole, posaconazole, anidulafungin, caspofungin, and micafungin was tested by the microdilution colorimetric method. A total of 364 C. parapsilosis complex isolates were identified by molecular methods: C. parapsilosis (90.7%), Candida orthopsilosis (8.2%), and Candida metapsilosis (1.1%). Most candidemias (C. parapsilosis, 76.4%; C. orthopsilosis, 70.0%; C. metapsilosis, 100%) were observed in adults. No C. orthopsilosis or C. metapsilosis candidemias occurred in neonates. C. parapsilosis was most frequent in adult intensive care unit (28.8%), surgery (20.9%), and internal medicine (19.7%) departments; and C. orthopsilosis was most frequent in hematology (28.6%), pediatrics (12.0%), and neonatology (11.5%) departments. The geographic distribution of C. orthopsilosis and C. metapsilosis was not uniform. According to CLSI clinical breakpoints, all C. orthopsilosis and C. metapsilosis isolates were susceptible to the nine agents tested. Resistance (MICs > 1 mg/liter) was observed only in C. parapsilosis: amphotericin B, posaconazole, itraconazole, and caspofungin (0.3% each), anidulafungin (1.9%), and micafungin (2.5%). Applying the new species-specific fluconazole and echinocandin breakpoints, the rates of resistance to fluconazole for C. parapsilosis and C. orthopsilosis increased to 4.8% and 0.3%, respectively; conversely, for C. parapsilosis they shifted from 1.9 to 0.6% (anidulafungin) and from 2.5 to 0.6% (micafungin). Our study confirms the different prevalence of C. parapsilosis complex candidemia among age groups: neither C. orthopsilosis nor C. metapsilosis was isolated from neonates; interestingly, C. metapsilosis was isolated only from adults and the elderly. The disparity in antifungal susceptibility among species could be important for therapy.

Introduction: Biofilm production is considered a potential virulence factor of some Candida species. For this reason, an understanding of biofilm behavior of Candida albicans and its closely related species Candida dubliniensis is key to the development of effective preventive measures for invasive and oral candidiasis. The aim of this study was to compare the capacity of biofilm production by blood and oral isolates of C. albicans and C. dubliniensis using polystyrene, flat-bottomed 100-well microtiter plates., Methods: A total of 47 isolates, consisting of 28 C. albicans (16 oral and 12 blood isolates) and 19 C. dubliniensis (11 oral and 8 blood isolates) were compared for their biofilm forming ability under aerobic and static conditions. XTT reduction assay was used to quantify the sessile growth., Results: All tested isolates produced biofilm, measured as XTT metabolic activity. Biofilm formation by C. albicans isolates was statistically significantly higher than biofilm formation by C. dubliniensis isolates at 24h (P=0.03) and 48 h (P=0.0001). There was a higher percentage (41.7%) of high producers of biofilms among C. albicans blood isolates than among oral isolates (31.3%), without statistically significant differences., Conclusions: This capability may allow C. albicans and C. dubliniensis to maintain their oral ecological niches as commensal or pathogenic microorganisms and can be a major virulence factor during invasive candidiasis. However, the differences in biofilm production among isolates should be taken into account when the anti-biofilm activity of antifungal agents or other virulence factors are tested in vitro., (Copyright © 2011 Elsevier España, S.L. All rights reserved.)

We have evaluated the in vitro activity of anidulafungin (AFG) against 31 strains of Candida parapsilosis sensu stricto by using broth microdilution, disk diffusion, and minimal fungicidal concentration (MFC) determination procedures. The two first methods showed a high level of activity of the drug, while MFCs were 1 to 5 dilutions higher than their corresponding MICs. To assess if MICs were predictive of in vivo outcomes, six strains representing different AFG MICs (0.12 to 2 μg/ml) were tested in a murine model of disseminated infection treated with different doses of the drug (1, 5, or 10 mg/kg of body weight). AFG was able to prolong the survival of mice infected with all the strains tested but was able to reduce the tissue burden of those mice infected only with the strains that showed the lowest MIC (0.12 μg/ml).

Genitourinary infections by Saccharomyces cerevisiae are rare. Here, we describe eight S. cerevisiae vulvovaginitis episodes where molecular (Affirm VPIII) and conventional microbiological methods (culture and carbohydrate assimilation) have proven to be inadequate for diagnostic purposes. DNA sequencing allowed the correct identification of the pathogen. All isolates were susceptible to most antifungal agents, with two of them also found to be susceptible-dose-dependent to itraconazole.

Invasive fungal diseases (IFD) are important causes of solid organ transplant-related morbidity and mortality. Modifications and improvements in the transplant surgical procedures, supportive care, and advances in the diagnosis and treatment of these IFD have produced notable changes in their epidemiology and outcome. Candida and other yeast genera continue to play an important etiological role, but Aspergillus and other filamentous fungi are the cause of most IFD in lung transplant recipients. This review is an update of the relevant findings in the literature related to the epidemiology, diagnosis and treatment of IFD in solid organ transplant recipients, with a main focus on invasive aspergillosis and candidiasis., (Copyright © 2011 Revista Iberoamericana de Micología. Published by Elsevier Espana. All rights reserved.)

Micafungin is a semisynthetic lipopeptide developed from Coleophoma empetri, which blocks the synthesis of β-1,3-D-glucan, an essential component of the fungal wall, though non-competitive inhibition of β-1,3-D-glucan synthetase. Micafungin is a dose-dependent candidacidal agent with excellent in vitro efficacy against most Candida spp. including species resistant to amphotericin B, such as Candida lusitaniae, several azoles, such as C. glabrata or C. krusei, and isolates not susceptible to other echinocandins. Moreover, this drug is active against Candida biofilms. Micafungin is a first-line drug for the treatment of candidemias and invasive candidiasis in adults and children (including neonates). This drug is approved for use in the treatment of invasive candidiasis and Candida esophagitis, as well as in the prophylaxis of Candida infections in hematopoietic stem cell transplant recipients or those at risk of prolonged neutropenia. Micafungin can be used both in the treatment and prevention of candidiasis in neonates, children, adolescents, adults, and the elderly, making it highly useful in patient groups in which the use of other antifungal drugs has not been authorized., (Copyright © 2011 Elsevier España S.L. All rights reserved.)

These guidelines are an update of recommendations for the diagnosis of invasive fungal infections by the Spanish Society of Clinical Microbiology and Infectious Diseases (SEIMC) published in 2004 (Enferm Infecc Microbiol Clin. 2004, 22:32-9). In this updated version of the guidelines, a comprehensive review of the most recent diagnostic innovations and levels of evidence to recommend those diagnostic procedures are included. We first analyse conventional diagnostic methods, microscopic examination and culture, underlining their limitations which have led to the development of alternative methods, such as fungal antigen and DNA quantification. Those alternative methods of diagnosis are analysed by fungal infection. We also briefly review the methods for molecular identification of fungal species and recommendations for carrying out susceptibility tests for antifungal drugs, including reference procedures, commercial techniques and their indications., (Copyright © 2010 Elsevier España, S.L. All rights reserved.)

Oral candidiasis is the most prevalent complication in HIV-infected and AIDS patients. Topical antifungal treatment is useful for the initial episodes of oral candidiasis, but most patients suffer more than one episode and fluconazole or itraconazole can help in the management, and voriconazole may represent a useful alternative agent for the treatment of recalcitrant oral and oesophageal candidiasis. The aim of this research was to study the in vitro activity of voriconazole and fluconazole against Mexican oral isolates of clinically relevant yeast. The in vitro susceptibility of 187 oral yeast isolates from HIV-infected and healthy Mexicans was determined for fluconazole and voriconazole by the M44-A disc diffusion method. At 24 h, fluconazole was active against 179 of 187 isolates (95.7 %). Moreover, a 100% susceptibility to voriconazole was observed. Voriconazole and fluconazole are highly active in vitro against oral yeast isolates. This study provides baseline data on susceptibilities to both antifungal agents in Mexico.

Candida albicans is the species most frequently isolated from oral specimens, but the recovery of other Candida species such as Candida dubliniensis is increasing. Differentiation of C. dubliniensis from C. albicans requires special tests and both species are misidentified in some studies. CHROM-Pal (CH-P) is a novel chromogenic medium used in our laboratory for differentiation between C. albicans and C. dubliniensis on the basis of colony colour and morphology, and chlamydospore production. The performance of CH-P and CHROMagar Candida (CAC) was compared for primary isolation and presumptive identification of yeasts from oral specimens from human immunodeficiency virus (HIV)-infected and uninfected individuals. The identification of Candida species on both media was compared with two reference identification methods (API ID 32 C and multiplex PCR). A total of 137/205 oral swabs (66.8 %) plated onto CH-P and CAC media were positive by culture and resulted in the growth of 171 isolates of Candida species on CH-P, whilst only 159 isolates grew on CAC. C. albicans was the most frequently isolated species in both groups of patients, followed by Candida parapsilosis in the HIV-negative group, and by C. dubliniensis in the HIV-infected group. The other Candida species isolated were Candida guilliermondii, Candida glabrata, Candida krusei, Candida tropicalis, Candida famata, Candida rugosa, Candida kefyr, Candida pelliculosa and Candida pulcherrima. The sensitivity and specificity for identifying C. albicans, C. krusei, C. tropicalis and C. dubliniensis on CH-P were over 98.5 %, always equal to or higher than those obtained when CAC was used. CH-P is a simple reliable medium for primary isolation and presumptive identification of yeast isolates from oral samples. The ability of CH-P to discriminate between C. dubliniensis and C. albicans was significantly higher (P <0.05) than that of CAC.

Background: The need to evaluate curricula for sponsorship for research projects or professional promotion, has led to the search for tools that allow an objective valuation. However, the total number papers published, or citations of articles of a particular author, or the impact factor of the Journal where they are published are inadequate indicators for the evaluation of the quality and productivity of researchers. The h index, proposed by Hirsch, categorises the papers according to the number of citations per article. This tool appears to lack the limitations of other bibliometric tools but is less useful for non English-speaking authors., Aims: To propose and debate the usefulness of the existing bibliometric indicators and tools for the evaluation and categorization of researchers and scientific journals., Methods: Search for papers on bibliometric tools., Results: There are some hot spots in the debate on the national and international evaluation of researchers' productivity and quality of scientific journals. Opinions on impact factors and h index have been discussed. The positive discrimination, using the Q value, is proposed as an alternative for the evaluation of Spanish and Iberoamerican researchers., Conclusions: It is very important de-mystify the importance of bibliometric indicators. The impact factor is useful for evaluating journals from the same scientific area but not for the evaluation of researchers' curricula. For the comparison of curricula from two or more researchers, we must use the h index or the proposed Q value. the latter allows positive discrimination of the task for Spanish and Iberoamerican researchers.

Background: Most recalcitrant infections are associated to colonization and microbial biofilm development. These biofilms are difficult to eliminate by the immune response mechanisms and the current antimicrobial therapy., Aim: To describe the antifungal of micafungin against fungal biofilms based in the scientific and medical literature of recent years., Methods: We have done a bibliographic retrieval using the scientific terms "micafungin", "activity", "biofilm", "Candida", "Aspergillus", "fungi", "mycos"*, susceptibility, in PubMed/Medline from the National Library of Medicine from 2006 to 2009., Results: Most current antifungal agents (amphotericin B and fluconazole) and the new azole antifungals have no activity against fungal biofilms. However, micafungin and the rest of echinocandins are very active against Candida albicans, Candida dubliniensis, Candida glabrata, and Candida krusei biofilms but their activities are variable and less strong against Candida tropicalis and Candida parapsilosis biofilms. Moreover, they have not activities against the biofilms of Cryptococcus y Trichosporon., Conclusions: The activity of micafungin against Candida biofilms gives more strength to its therapeutic indication for candidaemia and invasive candidiasis associated to catheter, prosthesis and other biomedical devices.

Background: Micafungin is a new and very useful pharmacological tool for the treatment of invasive mycoses with a wide antifungal spectrum for the most common pathogenic fungi. Micafungin is especially active against the genera Candida and Aspergillus. Its antifungal mechanism is based on the inhibition of the beta-1,3- D-glucan synthesis, an essential molecule for the cell wall architecture, with different con sequences for Candida and Aspergillus, being micafungin fungicide for the former and fungistatic for the latter., Aim: To describe the in vitro antifungal spectrum of micafungin based in the scientific and medical lite rature of recent years., Methods: We have done a bibliographic retrieval using the scientific terms, "micafungin", "activity", "Candida", "Aspergillus", "fungi", "mycos*", "susceptibility", in PubMed/Medline from the National Library of Medicine de EE.UU. from 2005 to 2009., Results: We can underline that most than 99% of Candida isolates are susceptible to < or = 2microg/ml of micafungin. MIC are very low (< or = 0.125microg/ml) for most clinical isolates of the species Candida albicans, Candida glabrata, Candida tropicalis and Candida krusei while Candida parapsilosis and Candida guilliermondii isolates are susceptible to anidulafungin concentrations < or = 2microg/ml. The activity of micafungin is excellent against those medical important species of Aspergillus. However, its activity is very low against Cryptococcus and the Zygomycetes., Conclusions: The excellent activity of micafungin has made this antifungal a first line therapeutic indication for candidemia and invasive candidiasis in non-neutropenic patients.

Anidulafungin is a new and very useful pharmacological tool for the treatment of invasive mycoses. The antifungal spectrum of anidulafungin reaches the most common pathogenic fungi. Anidulafungin is especially active against the genera Candida and Aspergillus. Its antifungal mechanism is based on the inhibition of the beta-1,3-D-glucan synthesis, an essential molecule for the cell wall architecture, with different consequences for Candida and Aspergillus, being anidulafungin fungicide for the former and fungistatic for the latter. This review describes the in vitro antifungal spectrum of anidulafungin based in the scientific and medical literature of recent years. We can underline that most than 99% of Candida isolates are susceptible to < or = 2 microg/ml of anidulafungin. MIC are very low (< or =0.125 microg/ml) for most clinical isolates of the species Candida albicans, Candida glabrata, Candida tropicalis and Candida krusei while Candida parapsilosis and Candida guilliermondii isolates are susceptible to anidulafungin concentrations < or = 2 microg/ml. An excellent activity of anidulafungin has been also described against Aspergillus, Pneumocystis and other fungi. However, its activity is very low against Cryptococcus and the Zygomycetes. The excellent activity of anidulafungin has made this antifungal a first line therapeutic indication for candidemia and invasive candidiasis in non-neutropenic patients.

Invasive infections caused by Candida spp. are increasing worldwide and are becoming an important cause of morbidity and mortality in immunocompromised patients. A large number of manifestations of candidiasis are associated with the formation of biofilms on inert or biological surfaces. Candida spp. biofilms are recalcitrant to treatment with conventional antifungal therapies. The aim of this study was dual 1) to determine the prevalence of biofilm producers among clinical isolates from catheter (16 C. albicans ) and blood culture (2 C. albicans and 30 C. tropicalis), and 2) to determine the activity of amphotericin B and anidulafungin against C. albicans and C. tropicalis biofilms of 24 and 48 hours of maturation. Biofilms were developed using a 96-well microtitre plate model and production and activity of antifungal agents against biofilms were determined by the tetrazolium (XTT) reduction assay. Of catheter and blood isolates, 62.5 and 56.25%, respectively, produced biofilms. By species, 68.42% of C. albicans and 53.33% of C. tropicalis were biofilm producers. C. albicans biofilms showed more resistance to amphotericin B and anidulafungin than their planktonic counterparts. Complete killing of biofilms was never achieved, even at the highest concentrations of the drugs tested. Anidulafungin displayed more activity than amphotericin B against C. albicans biofilms of 24 hours of maturation (GM MIC 0.354 vs. 0.686 microg/ml), but against C. tropicalis biofilms amphotericin B was more active (GM MIC 11.285 vs. 0.476 microg/ml). In contrast, against biofilms with 48 hours maturation, amphotericin B was more active against both species.

Objectives: Because of its considerable epidemiological relevance, accurate identification of Candida dubliniensis should be routinely performed in clinical microbiology laboratories. In an attempt to facilitate this task, the usefulness of the Bichro-Dubli test (Fumouze Diagnostics, Levallois-Perret, France) was assessed., Methods: Seventy-five collection strains (55 C. dubliniensis and 20 C. albicans) and 135 clinical yeast isolates that grew as green colonies in CHROMagar Candida were studied., Results: Bichro-Dubli was positive in 54 of 55 C. dubliniensis strains (sensitivity 98.2%) and negative in the 20 C. albicans strains (specificity 100%). The test identified 4 C. dubliniensis isolates among the 135 isolates cultured from clinical specimens., Conclusions: The Bichro-Dubli test is easy to perform and allows rapid identification of C. dubliniensis.

Voriconazole has been developed to meet the increasing need for new and useful antifungal agents for the treatment of invasive mycoses. This review describes the spectrum of voriconazole antifungal activity based on data from in vitro studies published during the last three years. This survey demonstrates that voriconazole has a broad antifungal spectrum against the most common fungal pathogens being its action fungistatic for Candida and fungicidal for Aspergillus and other filamentous fungi. Overall, more than 95% of all Candida isolates tested are susceptible to voriconazole and less than 3% are resistant. Similar or even better activity rates have been described for Aspergillus, Cryptococcus and most of yeasts and moulds of medical importance. We also discuss the limitations related to the azole cross-resistance observed in some Candida glabrata isolates, the poor activity of voriconazole against Scedosporium prolificans, its activity against fungal biofilms and the great potential usefulness of combination of voriconazole with other antifungal drugs.

The usefulness to diagnose and monitor invasive candidiasis (IC) using beta-glucan (BG) and antibodies against Candida albicans germ tubes (CAGT) was evaluated in a twice-weekly screening of 35 episodes in neutropenic adults at high risk. Three proven IC and three probable IC were assessed. Diagnostic levels of both markers were detected in 100% of proven IC and in 66% of probable IC. Sensitivity, specificity, positive and negative predictive values of BG and anti-CAGT antibodies detection were 83.3%, 89.6%, 62.5% and 96.3%, and 83.3%, 86.2%, 55.5%, 96.1%, respectively. False positive reactions occurred at a rate of 10.3% and 13.8% for the detection of BG and anti-CAGT antibodies, respectively. However, the patients with false positive results were different by each test. Both tests anticipated the clinical and radiological diagnosis, and the initiation of antifungal therapy in most patients. Combination of both tests improved specificity and positive predictive value to 100%.

Two cases of invasive candidiasis in intensive care patients are presented to illustrate the usefulness of detection of antibodies to Candida albicans germ tubes in the diagnosis of invasive candidiasis and in monitoring the efficacy of the antifungal treatment.

We have conducted a longitudinal study over a 3-year period to address the point prevalence, microbiological characteristics and antifungal susceptibility patterns of yeast isolates colonizing or infecting the oral cavities of 111 HIV-infected (51 adults, 60 children) and 201 non HIV-infected (109 adults, 92 children) Mexican persons. Regarding the epidemiology of oral candidiasis, Candida albicans was the most frequent species isolated. Seventy-one out of 85 isolates from colonized persons were C. albicans (83.5%), 27 isolates of them were from HIV-infected children and 44 from non HIV-infected patients. Sixty-two isolates belonged to serotype A which was the most prevalent serotype of C. albicans. Non-albicans species (Candida glabrata, Candida tropicalis and Candida parapsilosis, and Saccharomyces cerevisiae) were isolated from 16.5% of colonized patients and from 38.5% patients with candidiasis or Candida-related lesions. There were nine episodes of infection or colonization by at least 2 different yeast species. In the case of HIV/AIDS patients, it was determined that yeast carriage was not associated with the number of CD4+ cells or the viral load, but HAART reduced the prevalence of oral candidiasis. Overall, most patients harbored strains in vitro susceptible to fluconazole, however 10.8% of the yeasts were resistant to one or more azole antifungal agents and 29% were intermediate susceptible to them. On the contrary, 5-fluorocytosine was very active against all isolates tested, and amphotericin B was active against 97.9% of them.

We have studied the in vitro antifungal activity of voriconazole, fluconazole and itraconazole against 252 clinical isolates of dermatophytes and Scopulariopsis brevicaulis by a standardized agar diffusion method (NeoSensitabs). Several important factors such as temperature (28 degrees C vs. 35 degrees C) and incubation time (2-10 days vs. 18-74 h) were adapted to dermatophytes and Scopulariopsis requirements. Voriconazole showed an excellent activity against most species of dermatophytes, higher than itraconazole and fluconazole. However, S. brevicaulis isolates were highly resistant to all azoles used in this study. Voriconazole might be an interesting antifungal alternative to refractory superficial mycoses.

Background: Denture stomatitis is associated to Candida albicans, different bacteria and other co-factors such as an acid pH, a carbohydrate ingestion increase, different systemic illnesses and pharmacological treatments., Objective: The aim of this study was to determine Candida albicans, Staphylococcus aureus and Streptococcus mutans prevalence in the mucous membrane and prosthesis of patients with and without atrophic denture stomatitis and its relationship with other potential clinical co-factors., Study Design: Saliva was collected from 105 patients (62 female and 43 male) wearing dental prosthesis in order to measure their pH. Oral samples of the mucous membrane and the internal surface of dental prosthesis were taken with sterile cotton to proceed with the microbiological study. The identification of the isolated microorganisms was performed using conventional microbiological methods., Results: Diabetes and Hypertension were the most frequent systemic illnesses. High carbohydrate ingestion was observed in numerous patients. Atrophic denture stomatitis was reported in 50 patients and the pH average in saliva was of 5.2. The presence of C albicans, S. aureus and S. mutans in the mucous membrane and prosthesis was of 51.4%, 52.4% and 67.6%, respectively. C. albicans was isolated in 66.7% from the prosthesis, whereas S. aureus and S. mutans were isolated in 49.5% of those same prosthesis. C. albicans was isolated in 86% of the patients with atrophic denture stomatitis and S. aureus was isolated in a similar percentage (84% of patients). The isolation of S. mutans was less frequent, and it was observed in 16% of the oral samples of these patients., Conclusions: C. albicans, S. aureus and S. mutans frequently colonize the oral mucous of patients wearing dental prosthesis. This illness-bearing condition is more frequent in patients with denture stomatitis, even though dental prosthesis colonization is lower than in the oral mucous.

Melanin production is one of the most important criteria for rapid identification of Cryptococcus neoformans. Most of the media described in the literature for identifying C. neoformans are very complex; they contain many organic or inorganic compounds and are difficult to prepare and store. The new minimal synthetic caffeic acid medium described in this paper is simpler to prepare, convenient and constitutes an interesting new medium for the rapid identification of C. neoformans isolates.

The aim of this study was to determine the prevalence of primary resistance to 5-fluorocytosine (5FC) among clinical isolates of yeasts in Spain where this drug is not currently available for therapy. We have tested the in vitro activity of 5FC against 1,021 recent yeast clinical isolates, including 522 Candida albicans, 140 Candida parapsilosis, 68 Candida glabrata, 41 Candida dubliniensis, 50 Candida guilliermondii, 34 Candida tropicalis, 28 Candida krusei, 20 Candida famata, 11 Cryptococcus neoformans, 5 Cryptococcus albidus, 43 Rhodotorula spp., 24 Trichosporon spp., 5 Saccharomyces cerevisiae, 9 Pichia spp., and 21 isolates from other 11 yeast species. The MICs were determined by the ATB Fungus agar microdilution test (bioMerieux, France) and the following interpretive breakpoints were used: susceptible, > 4 microg/ml; intermediate, 8 to 16 microg/ml; resistant, > 32 microg/ml. 5FC was very active against Candida spp. and other medically important yeasts as 852 (83.4%) of the studied isolates were susceptible (MIC < 4 microg/ml). The species most susceptible to 5FC were C. dubliniensis (100%of isolates; MIC90, 0.25 microg/ml), C. famata (100% of isolates; MIC90, 0.25 microg/ml), C. guilliermondii (98%of isolates; MIC90, 0.25 microg/ml), C. glabrata (95.5% of isolates; MIC90, 0.25 microg/ml), and C. neoformans (90.9% of isolates; MIC90, 2 microg/ml). Primary resistance to 5FC was very uncommon, and a MIC > 32 microg/ml, indicator of in vitro resistance, was observed in 106 isolates (10.4%): 77 C. albicans (16.5% of isolates; MIC90, > 128 microg/ml), 9 C. parapsilosis (6.4% of isolates; MIC90, 8 microg/ml), 4 C. albidus (80% of isolates, MIC50, > 128 microg/ml), 3 C. glabrata (4.4% of isolates; MIC90, 0.25 microg/ml), 3 C. tropicalis (8.8% of isolates; MIC90, 4 microg/ml), 2 C. krusei (7.1% of isolates; MIC90, 8 microg/ml), 2 Rhodotorula spp. (4.6% of isolates, MIC90, 1 microg/ml), 8 Trichosporon spp. (33.3% of isolates; MIC90, 64 microg/ml), and 1 C. lipolytica (50% of isolates). Interestingly, most C. albicans (67 out of 77 isolates) resistant to 5FC were serotype B isolates.

Twenty-six Candida dubliniensis and 27 Candida albicans oral strains isolated from patients infected by the human immunodeficiency virus (HIV) were tested for germ tube production and 21 extracellular enzymatic activities. Assessment of the enzymatic profile was performed by using the API-ZYM commercial kit system (bioMerieux, France), which tests 19 different enzymes. Protease activity was expressed during the first days of incubation by 100% of the strains studied and resulted higher than phospholipase activity in the C. dubliniensis and C. albicans strains tested. The API-ZYM profile of the C. dubliniensis and C. albicans strains differs with respect to the number and percentage of the enzymes considered, as well as with the intensity of the substrate metabolized by the strains, in particular for the enzymes n 8 (cystine-arylamidase), n 12 (naphtol-AS-BI-phosphohydrolase) and n 16 (alpha-glucosidase). These enzymes may be useful to differentiate C. dubliniensis and C. albicans together with other phenotypic characteristics proposed in the literature. No relationship among protease, phospholipase and other extracellular enzymatic activities was observed in C. dubliniensis. The average percentage of strains filamentation after 4 h was between 32 and 42%.

During the last decades, the use of antibody tests for the diagnosis of invasive mycoses has declined as a consequence of the general belief that they are insensitive and non-specific. However, there is a clear evidence that antibodies can be detected in highly immunodeficient patients (such as bone marrow transplant recipients), and that those antibodies are useful for the diagnosis. Antibody tests are currently in use as diagnostic tools for some primary mycoses, such as the endemic mycoses, aspergilloma, allergic bronchopulmonary aspergilosis and sporothrichosis. For invasive candidiasis, diagnostic methods must differentiate Candida colonization of mucous membranes or superficial infection from tissue invasion by this microorganism. Substantial progress has been made in diagnosis of invasive candidiasis with the development of a variety of methods for the detection of antibodies and antigens. However, no single test has found widespread clinical use and there is a consensus that diagnosis based on a single specimen lacks sensitivity. It is necessary to test sequential samples taken while the patient is at greatest risk for developing invasive candidiasis to optimize the diagnosis. Results obtained from a panel of diagnostic tests in association with clinical aspects will likely be the most useful strategy for early diagnosis and therapy.

Introduction: Two tests for the detection of antibodies to Candida albicans germ tubes in patients with invasive candidiasis were compared: a new commercially available test (Candida albicans IFA IgG) and the indirect immunofluorescence test generally used for this purpose., Methods: With the use of two indirect immunofluorescence tests, retrospective study was done on 172 sera from 51 patients classified into two groups: Group I included 123 serum samples from 32 patients with invasive candidiasis, and Group II, the control, included 49 serum samples from 19 patients with no evidence of Candida infection., Results: In Group I, 84% of patients presented anti-germ tube antibody titers >or= 1:160 by the Candida albicans IFA IgG test and 78.1% of patients were positive by the generally used test. There was a high correlation between the two tests (R2 =0.9512 by patients; R2 = 0.8986 by sera). When a titer value of >or= 1:160 was used as cutoff, the Candida albicans IFA IgG test showed a sensitivity of 84.4% and a specificity of 94.7%, whereas the traditional test showed a sensitivity of 78.1% and a specificity of 100%., Conclusions: The commercially available Candida albicans IFA IgG test is similar to the test generally used for the detection of antibodies to C. albicans germ tubes and provides faster and easier diagnosis of invasive candidiasis in the clinical microbiology laboratory.

Infections caused by fungi (mycoses) are increasingly reported in many countries owing to greater life expectancy associated with an increase in quality of medical and surgical procedures, as well as the emergence of diseases or infections that affect the immune system such as AIDS. Nosocomial outbreaks of fungal infections are sometimes reported, and typing is then necessary to find the reservoirs, analyze the modes of transmission, study the antifungal susceptibility patterns, and investigate the susceptibility of the host. In addition, the food industry is increasingly demanding typing methods that could help in selection of the best fungal strains, in order to incorporate them in the productive chains and augment the quality and security of food. This is the case for Saccharomyces cerevisiae in the wine industry: the selection and characterization of indigenous or autochthonous strains is an important objective for the production of high-quality certified wines.Several genotyping methods are now widely used for strain delineation of medically or economically important microorganisms belonging to the kingdom Fungi. Most molecular typing methods are comparable to those already described for bacteria, although the peculiarities of their nucleic acids increase the number of available methods. Although typing procedures based on the analysis of nucleic acid sequences have been developed, most genotyping methods currently in use are electrophoretically based, and the procedures include the visual comparison of nucleic acid band profiles or their reading with the help of computerized software. Here we describe some of the most frequently used genotyping methods for fungi, based on polymerase chain reactions (PCR), the isolation of chromosomal or mitochondrial DNA, and their restriction using endonuclease enzymes. The latter methods are exclusive for typing eukaryotic organisms and are based on the expected polymorphism obtained from the separation of large chromosomes using pulsed-field gel electrophoresis (PFGE) and the restriction of mitochondrial or chromosomal DNA. More sophisticated methods, such as those that combine endonuclease restriction with hybridization, are also available, although their use is less extensive and is limited mostly to research laboratories.

Twenty-seven Candida albicans strains and 26 Candida dubliniensis strains, isolated from HIV patients, were tested for their adherence to buccal and vaginal epithelial cells. Both species showed important levels of adhesion to buccal and vaginal epithelial cells, although C. albicans showed the highest levels of adhesion. These results suggest that both Candida species are well adapted, in terms of adhesion capability, to the oral and vaginal environment.

The ability of Candida glabrata to switch between different colony phenotypes on a Sabouraud's-chloramphenicol agar supplemented with phloxine B was assessed in 14 C. glabrata isolates. Three phenotypes (pale pink smooth, dark pink smooth and fuchsia petite) were observed in vitro and two of them (pale pink smooth and dark pink smooth) in fresh clinical isolates plated directly from vaginal specimens from patients with C. glabrata vaginitis and patients colonized with C. glabrata. The pale pink smooth phenotype was the predominant and the remaining phenotypes can be derived from it. No changes in susceptibility to different antifungals were observed in the pale pink smooth, dark pink smooth, fuchsia petite phenotypes but they showed differences in cell wall antigens.