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PubMed1996

Evaluation of a commercial medium for identification of Candida species.

CHROMagar Candida (CHROMagar, France) was evaluated as a medium for the presumptive identification and isolation of yeasts using 1,537 isolates of medically important yeasts, including 970 Candida albicans, 165 Candida parapsilosis, 131 Candida glabrata, 62 Candida guilliermondii, 35 Candida krusei, 32 Candida tropicalis, 31 Rhodotorula rubra, 23 Trichosporon spp. (17 Trichosporon beigelii), 17 Candida famata, 16 Candida pelliculosa, 10 Pichia etchelsii, 10 Saccharomyces cerevisiae, 8 Candida lusitaniae, 7 Cryptococcus spp., and 20 isolates of other Candida spp. After 48 h of incubation at 37 degrees C, the sensitivity and specificity were, respectively, 99% and 100% for Candida albicans, 93.8% and 99.1% for Candida tropicalis, and 100% and 100% for Candida krusei. In addition to colony color, other colony characteristics were important for identification of some species, such as rough colonies in Candida krusei isolates or the halo around the colonies of Candida tropicalis. A great variety of colors was observed among species other than Candida albicans, Candida tropicalis, and Candida krusei. For identification purposes, CHROMagar Candida medium has an accuracy similar to that of germ-tube tests and chlamydospore development tests for Candida albicans and to that of the ATB ID32C kit (API, bioMérieux, France) for Candida tropicalis and Candida krusei.

Candida↗
PubMed2002

New chromogenic agar medium for the identification of Candida spp.

A new chromogenic agar medium (Candida diagnostic agar [CDA]) for differentiation of Candida spp. is described. This medium is based on Sabouraud dextrose agar (Oxoid CM41) and contains (per liter) 40.0 g of glucose, 10.0 g of mycological peptone, and 15.0 g of agar along with a novel chromogenic glucosaminidase substrate, ammonium 4-(2-[4-(2-acetamido-2-deoxy-beta-D-glucopyranosyloxy)-3-methoxyphenyl]-vinyl)-1-(propan-3-yl-oate)-quinolium bromide (0.32 g liter(-1)). The glucosaminidase substrate in CDA was hydrolyzed by Candida albicans and Candida dubliniensis, yielding white colonies with deep-red spots on a yellow transparent background after 24 to 48 h of incubation at 37 degrees C. Colonies of Candida tropicalis and Candida kefyr were uniformly pink, and colonies of other Candida spp., including Candida glabrata and Candida parapsilosis, were white. CDA was evaluated by using 115 test strains of Candida spp. and other clinically important yeasts and was compared with two commercially available chromogenic agars (Candida ID agar [bioMerieux] and CHROMagar Candida [CHROMagar Company Ltd.]). On all three agars, colonies of C. albicans were not distinguished from colonies of C. dubliniensis. However, for the group containing C. albicans plus C. dubliniensis, both the sensitivity and the specificity of detection when CDA was used were 100%, compared with values of 97.6 and 100%, respectively, with CHROMagar Candida and 100 and 96.8%, respectively, with Candida ID agar. In addition, for the group containing C. tropicalis plus C. kefyr, the sensitivity and specificity of detection when CDA was used were also 100%, compared with 72.7 and 98.1%, respectively, with CHROMagar Candida. Candida ID agar did not differentiate C. tropicalis and C. kefyr strains but did differentiate members of a broader group (C. tropicalis, C. kefyr, Candida lusitaniae plus Candida guilliermondii); the sensitivity and specificity of detection for members of this group were 94.7 and 93.8%, respectively. In addition to the increased sensitivity and/or specificity of Candida detection when CDA was used, differentiation of colony types on CDA (red spotted, pink, or no color) was unambiguous and did not require precise assessment of colony color.

Agar↗
PubMed1978

Damage to pseudohyphal forms of Candida albicans by neutrophils in the absence of serum in vitro.

Large forms of Candida are characteristically present in invasive lesions and are often cleared by host defenses. Therefore, an in vitro system was developed to study interactions between leukocytes and pseudohyphae. By light, phase contrast, and electron microscopic observations, in the absence of serum, neutrophils attached to and spread over the surfaces of partially ingested pseudohyphae, which then appeared damaged. Using a new assay which measured neutrophil-induced inhibition of uptake of [(14)C]cytosine by Candida, damage to Candida in the absence of serum was 53.04+/-2.96% by neutrophils from 27 normal subjects. With serum, damage to Candida increased because of opsonization by low levels of anti-Candida immunoglobulin G in normal sera. Damage to Candida was inhibited by colchicine, cytochalasin B, and 2-deoxyglucose, which interfered with spreading of neutrophils over the surfaces of Candida. Dibutyryl cyclic AMP, theophylline, and isoproterenol also inhibited damage to Candida. Hydrocortisone was inhibitory in levels (10 muM) achievable with pharmacologic doses in man. Light, fluorescence, and electron microscopy indicated that neutrophils degranulated after contact with Candida. Quantitative studies revealed only a minimal increase in specific release of lysosomal enzymes from azurophil granules, but much greater release of lysozyme from specific granules. Candida activated neutrophil oxidative microbicidal mechanisms, as shown by iodination of Candida by neutrophils, and chemiluminescence from neutrophils interacting with Candida. Unlike live Candida, killed Candida did not induce chemiluminescence, were not iodinated, and did not attach to neutrophils by microscopy. Like Candida pseudohyphae, contact between neutrophils and hyphal forms of Aspergillus and Rhizopus occurred in the absence of serum. This did not occur with Cryptococcus neoformans, an encapsulated yeast, and was low with Candida yeasts. These findings indicate that neutrophils can recognize and attach to Candida pseudohyphae, then damage the Candida. This may represent a general reaction between neutrophils and large forms of fungi. Though the size of the organisms precludes complete ingestion, neutrophil oxidative microbicidal mechanisms are activated, and preferential release of contents of specific granules appears to occur.

Antibodies, Fungal↗
PubMed2002

Systemic Candida infection in burn patients: a case-control study of management patterns and outcomes.

BACKGROUND: Burn patients are ideal hosts for opportunistic infections. Candida infection in burn patients has a reported mortality ranging from 14% to 90%. This retrospective case-control study compares management patterns and outcomes of burn patients who develop systemic Candida with those who do not. METHODS: Inpatients at our burn center with two or more positive culture sites for Candida from January 1, 1995, through December 31, 2000 and who sustained burn injury of >/=10% total body surface area (TBSA) were identified. A cohort of patients without Candida was matched for age and size of burn injury using our institution's TRACS/ABA trade mark registry. Management variables included days to burn wound coverage; use of artificial dermis; number of antibiotic days; treatment with imipenem, vancomycin, or aminoglycosides; need for abdominal surgery; and receipt of tracheostomy. Outcome measures were hospital length of stay (LOS) and mortality. RESULTS: Candida patients (n = 44) had a mean age of 39.8 years, and sustained an average burn size of 47.2% TBSA with 28.6% full-thickness injury. Controls (n = 44) had a mean age of 39.8 years, and sustained an average burn size of 46.0% TBSA with 26.6% full-thickness injury. Patients with multiple Candida sites required 36 days to achieve burn wound coverage with autograft versus 21 days for the control group (P = 0.004). Candida patients were significantly more likely to be managed with artificial dermis than were controls (Odds Ratio = 9.56, 95% Confidence Interval = 1.64-181.53). Patients with Candida infection averaged 72 days of treatment with systemic antibiotics, whereas the controls averaged only 36 days of antibiotic treatment (P = 0.001). Further, patients with multiple sites of Candida were more likely to have received imipenem, vancomycin, or an aminoglycoside (Odds Ratio = 11.99, 95% Confidence Interval = 3.10-79.71). Mean LOS was 62 days for patients with Candida and 30 days for the controls (P < 0.001). The mortality rate in patients with Candida was 23%, which did not differ significantly from the 27% mortality rate of the controls. CONCLUSION: Early wound coverage with autograft clearly decreases the likelihood of systemic Candida infection in burn patients. Patients who received artificial dermis as a component of their wound management strategy more often developed systemic Candida infection in this series. Burn patients who require prolonged courses of antibiotics or treatment with broad-spectrum antibiotics should be carefully monitored for the development of Candida. Survival of burn patients who develop systemic Candida infection is no different from survival in comparable burn patients who do not acquire Candida. Future research should address optimal management of Candida infection in burn patients.

Adult↗
PubMed1994

An experimental model for study of Candida survival and transmission in human volunteers.

In order to determine the potential for cross-transmission of Candida spp. between health-care workers and patients, the survival of clinical isolates of five species of Candida on the palms of human volunteers was tested. One hundred microliters of a McFarland 1.0 density suspension (5 x 10(5) cfu) from an overnight culture of Candida albicans, Candida krusei, Candida parapsilosis, Candida tropicalis and Candida glabrata was used as inoculum. The degree of hydrophobicity of the different Candida species was also tested and did not influence the survival. The half-lives were brief, being 9.5, 12.4, 7.4, 12.8, 9.6 min for Candida albicans, Candida krusei, Candida glabrata, Candida parapsilosis, and Candida tropicalis, respectively, but at 45 min 2.6 x 10(3) to 3 x 10(4) organisms remained on the hands. Survival of Candida albicans for as long as 24 h on inanimate surfaces was observed. Transmission from one hand to a second hand occurred in 69% of the experiments and from the first to a third hand in 38%. Transmission to and from inanimate surfaces was successful in most of the experiments (90%). This experimental model aids in the biological study of Candida spp. and suggests some of the potential mechanisms of transmission.

Candida↗
PubMed1994

Detection of Candida casts in experimental renal candidiasis: implications for the diagnosis and pathogenesis of upper urinary tract infection.

The distinction between upper versus lower urinary tract infection in patients with candiduria is a commonly encountered and therapeutically important diagnostic dilemma. Candida casts have been reported in the urine of several individual case reports of human renal candidiasis. The specificity of Candida casts would identify unequivocally a patient with upper urinary tract disease. Little is known, however, about the sensitivity and the formation of Candida casts. We therefore studied the diagnostic yield, methods for detection and pathogenesis of Candida cast formation in serially collected urine specimens from immunologically intact and granulocytopenic rabbit models of haematogenous disseminated candidiasis. Refractile blastoconidia and pseudohyphae of Candida encased in the granular matrix were seen on wet mounts while Candida stained a brilliant red in the fuschia pink tubular matrix on periodic acid Schiff (PAS) stained cytopathology filters. Among 24 rabbits with disseminated candidiasis, 11 (46%) had Candida casts detectable by wet mount and PAS-stained urine filters in comparison to none of 10 non-infected immunologically normal controls (P = 0.014). Fifteen (70%) of 21 episodes of Candida casts were detected within the first 3 days of infection, indicating possible utility in the early diagnosis of renal candidiasis. No Candida casts were detected in the urine of granulocytopenic rabbits, possibly due to the rapid destruction of tubules and abrogation of cast formation. This absence of detectable Candida in eight infected granulocytopenic rabbits differed significantly from that of 24 non-granulocytopenic infected rabbits, in which Candida casts were detected in 11 (46%) (P = 0.029). Candida cast formation occurred predominantly in the cortex. Histopathological examination demonstrated invasion of Candida into the glomerular tufts and peritubular capillaries, followed by development of Candida casts in the proximal and distal tubules, respectively. Detection of renal Candida casts may be a useful diagnostic marker in distinguishing upper versus lower urinary tract candidiasis.

Animals↗
PubMed1995

Differential susceptibility of yeast and hyphal forms of Candida albicans to macrophage-derived nitrogen-containing compounds.

Candida albicans is a dimorphic fungus capable of transition from the yeast form (Y-Candida) to the hyphal form (H-Candida). Both Y-Candida and H-Candida are known to be growth inhibited by murine macrophages (M phi) in vitro. In the present report, we demonstrate that M phi exposed to interferon gamma (IFN-gamma) plus lipopolysaccharide (LPS) show enhanced anti-Y-Candida and anti-H-Candida activities. To further investigate the phenomenon, Y-Candida and H-Candida were assessed for susceptibilities to M phi-derived supernatants. Only the growth of H-Candida, and not that of Y-Candida, is impaired by cell-free supernatants from M phi treated with IFN-gamma plus LPS. In contrast, no H-Candida growth inhibition occurs when supernatants from M phi exposed to IFN-gamma plus LPS in the presence of NG-monomethyl-L-arginine, an inhibitor of nitric oxide (NO) synthesis, are employed. Finally, supernatants from M phi incubated with sodium nitroprusside, an NO-generating agent, also show anti-H-Candida activity. In conclusion, these results indicate that H-Candida but not Y-Candida is susceptible to extracellular antifungal mechanisms employed by M phi, which likely involve stable nitrogen-containing compounds.

Animals↗
PubMed2001

Candida albicans is phagocytosed, killed, and processed for antigen presentation by human dendritic cells.

Candida albicans is a component of the normal flora of the alimentary tract and also is found on the mucocutaneous membranes of the healthy host. Candida is the leading cause of invasive fungal disease in premature infants, diabetics, and surgical patients, and of oropharyngeal disease in AIDS patients. As the induction of cell-mediated immunity to Candida is of critical importance in host defense, we sought to determine whether human dendritic cells (DC) could phagocytose and degrade Candida and subsequently present Candida antigens to T cells. Immature DC obtained by culture of human monocytes in the presence of granulocyte-macrophage colony-stimulating factor and interleukin-4 phagocytosed unopsonized Candida in a time-dependent manner, and phagocytosis was not enhanced by opsonization of Candida in serum. Like macrophages (Mphi), DC recognized Candida by the mannose-fucose receptor. Upon ingestion, DC killed Candida as efficiently as human Mphi, and fungicidal activity was not enhanced by the presence of fresh serum. Although phagocytosis of Candida by DC stimulated the production of superoxide anion, inhibitors of the respiratory burst (or NO production) did not inhibit killing of Candida, even when phagocytosis was blocked by preincubation of DC with cytochalasin D. Further, although apparently only modest phagolysosomal fusion occurred upon DC phagocytosis of Candida, killing of Candida under anaerobic conditions was almost equivalent to killing under aerobic conditions. Finally, DC stimulated Candida-specific lymphocyte proliferation in a concentration-dependent manner after phagocytosis of both viable and heat-killed Candida cells. These data suggest that, in vivo, such interactions between DC and C. albicans may facilitate the induction of cell-mediated immunity.

Antigen Presentation↗
PubMed2006

Global surveillance of in vitro activity of micafungin against Candida: a comparison with caspofungin by CLSI-recommended methods.

Micafungin is an echinocandin antifungal agent that has recently been approved for the prevention of invasive fungal infection and the treatment of esophageal candidiasis. Prospective sentinel surveillance for the emergence of in vitro resistance to micafungin among invasive Candida sp. isolates is indicated. We determined the in vitro activity of micafungin against 2,656 invasive (bloodstream or sterile site) unique patient isolates of Candida spp. collected from 60 medical centers worldwide in 2004 and 2005. We performed antifungal susceptibility testing according to the Clinical and Laboratory Standards Institute (CLSI) M27-A2 method and used a 24-hour prominent inhibition endpoint for determination of the MIC. Caspofungin was tested in parallel against all isolates. Of 2,656 invasive Candida sp. isolates, species distribution was 55.6% Candida albicans, 14.4% Candida parapsilosis, 13.4% Candida glabrata, 10.1% Candida tropicalis, 2.4% Candida krusei, 1.7% Candida guilliermondii, 0.9% Candida lusitaniae, 0.6% Candida kefyr, and 0.9% other Candida species. Overall, micafungin was very active against Candida (MIC50/MIC at which 90% of the isolates tested are inhibited [MIC90], 0.015/1 microg/ml; 96% inhibited at a MIC of < or =1 microg/ml, 100% inhibited at a MIC of < or =2 microg/ml) and comparable to caspofungin (MIC50/MIC90, 0.03/0.25 mug/ml; 99% inhibited at a MIC of < or =2 microg/ml). Results by species, expressed as MIC50/MIC90 (micrograms per milliliter), were as follows: C. albicans, 0.015/0.03; C. glabrata, 0.015/0.015; C. tropicalis, 0.03/0.06; C. krusei, 0.06/0.12; C. kefyr, 0.06/0.06; C. parapsilosis, 1/2; C. guilliermondii, 0.5/1; C. lusitaniae, 0.12/0.25; other Candida spp., 0.25/1. Although the species distribution varied considerably among the different geographic regions, there was no difference in micafungin activity across the regions. Micafungin has excellent in vitro activity against invasive clinical isolates of Candida from centers worldwide.

Antifungal Agents↗
PubMed2003

Candidemia and the susceptibility pattern of Candida isolates in blood.

OBJECTIVE: Candida species has become one of the most common blood isolates as well as one of the leading causes of nosocomial bloodstream infections. The purpose of our study was to determine the prevalence of Candida species among our bloodstream infecting organisms and the susceptibility pattern of the Candida isolates to antifungal agents. METHODS: A prospective study was carried out in the Division of Microbiology, King Khalid National Guard Hospital, Jeddah, Kingdom of Saudi Arabia of all positive blood cultures for Candida species. The study took place from 1st January 1998 to March 2002. Identification and susceptibility pattern of isolates were determined by the Candifast technique to amphotericin B, fluconazole, nystatin, Flucytosine, econazole, ketoconazole and miconazole. RESULTS: Over a 2-year period, 17,916 blood cultures were performed in our hospital. There were 2,972 positive cultures, of which 83 (2.8%) patients had Candida species isolated from their bloodstream. Of these, 38 (46%) were Candida albicans (C.albicans). The remaining 45 strains were made up of Candida tropicalis 9 (10.8%); Candida parapsilosis 9 (10.8%); Candida species 9 (10.8%); Candida guilliermondi 6 (7.2%); Candida krusei 5 (6%); Candida glabrata 4 (4.8%); Candida pseudotropicalis 2 (2.4%) and Trichosporon species 1 (1.2%). All Candida species were susceptible to amphotericin B. However, only 18 (47%) out of 38 C.albicans were susceptible to fluconazole, while only 8 (17.7%) of 45 non-C.albicans strains were susceptible to this drug. CONCLUSION: The susceptibility of C.albicans to fluconazole in our hospital using the Candifast method is very low (47%). These results need to be confirmed by carrying out the Etest or the NCCLS M27-A method to confirm the true susceptibilities of Candida strains in our locality.

Antifungal Agents↗
PubMed2000

Determinants of different Candida species infections of the genital tract in women. Sporachrom Study Geoup.

OBJECTIVE: We have analyzed the differences in the epidemiological characteristics of women with different Candida low female genital tract infection. STUDY DESIGN: Eligible for the study were 4228 women aged 18-70 years with symptomatic low gynecological tract infection and clinical findings suggestive for Candida infection consecutively attending during the study period first level outpatients gynecological services in Italy. CHROMagar Candida method was used to identify albicans and non-albicans species and among non-albicans ones Candida glabrata, tropicalis and krusei. RESULTS: Out of the 4228 women who entered the study, Candida infection was confirmed by CHROMagar test in 3351 cases (79.3%): Candida albicans was identified in 1431 cases (43%) and non-albicans in 1920. Among the 1920 women with non-albicans infection, Candida glabrata was identified in 1207 women, Candida krusei in 290, Candida tropicalis in 404 (in 19 cases other species or non-specified species were involved). Candida albicans infection was more frequently reported than non-albicans ones in diabetic women (Odds Ratio, OR=1.7, 95%, Confidence Interval, CI 1.1-2.7). Current oral contraceptive users tended more frequently to be infected with Candida albicans than non-albicans, however the estimated OR was only slightly above unity and of borderline statistical significance (OR 1.3, 9.5%, CI 1.1-1.5). Women reporting previous treatment with topic antimicotic reported more frequently non-albicans infection, than Candida albicans ones. However the association was limited and of borderline statistical significance (OR albicans vs. non albicans 0.7, 95% CI 0.5-1.0). Albicans infection was more frequently identified in women whose partner reported symptomatology for Candida infection (OR 1.7, 95% C.I. 1.4-2.0). CONCLUSIONS: This study shows that in this Italian population with symptomatic Candida infection of low female genital tract, there are some differences in the epidemiological characteristics of women with albicans and non-albicans infection.

Adolescent↗
PubMed1997

Significance of the isolation of Candida species from respiratory samples in critically ill, non-neutropenic patients. An immediate postmortem histologic study.

The diagnosis of pulmonary candidiasis is still controversial. We undertook a prospective study on 25 non-neutropenic, mechanically ventilated (> 72 h) patients who died in our ICU with the aim of assessing the incidence and significance of the isolation of Candida species from quantitative cultures of immediate postmortem lung biopsies and different respiratory sampling techniques. Immediate postmortem respiratory samples (endotracheal aspirate, protected specimen brush [PSB], bronchoalveolar lavage [BAL], blind biopsies [average 14/patient], and bilateral bronchoscopically guided biopsies [two per patient]) were taken from all patients. Lung tissue specimens were histologically examined. Respiratory samples were classified as having Candida or otherwise. Ten (40%) patients had at least one pulmonary biopsy yielding Candida spp. Among these 10 patients with Candida isolates, only two had definite pulmonary candidiasis. A total of 470 microorganisms were isolated from 280 of 375 (77%) lung biopsy samples in all 25 patients. Candida species represented 9% (n = 40) of the isolates, corresponding to 10 patients (40%). In the 10 patients in whom Candida species was isolated from pulmonary biopsies, this was always associated with the isolation of the same microorganism from one of the sampling methods. Quantitative cultures of Candida species from different sampling methods correlated well among each other but could not discriminate the presence from absence of Candida pneumonia. A logistic regression model adjusted for the presence of antibiotics, days of antibiotic treatment, mechanical ventilation period, age, ARDS, parenteral nutrition, and gender did not show any independent risk factor for developing positive pulmonary samples for Candida species. The incidence of Candida isolation from pulmonary biopsies in critically ill mechanically ventilated, non-neutropenic patients who die is high (40%). However, the incidence of definite Candida pneumonia was 8%. We also found that Candida colonization is uniform throughout the different lung regions, and that the presence of Candida in respiratory samples, independently of quantitative cultures, is not a good marker of Candida pneumonia in critically ill, non-neutropenic, non-AIDS patients.

Adult↗
PubMed2001

Molecular epidemiology of Candida colonizing critically ill patients in intensive care units.

BACKGROUND AND PURPOSE: Nosocomial Candida infections are an important cause of morbidity and mortality in critically ill patients. Although there is growing evidence that candidemia develops primarily as a consequence of endogenous colonization, hospital outbreaks of Candida infection are not uncommon. To examine the prevalence and consequence of Candida colonization in critically ill patients and to elucidate the contribution of cross-transmission to the high frequency of nosocomial fungal infection in intensive care units (ICUs), a 6-month prospective surveillance study was conducted. PATIENTS AND METHODS: A total of 342 adult patients with an expected ICU stay of 48 hours or more were enrolled in the study. Surveillance cultures were taken from the rectal region, oropharynx, and urine on ICU entry and weekly thereafter. The electrophoretic karyotypes (EKs) of all isolates were characterized using pulsed-field gel electrophoresis. RESULTS: A total of 873 Candida strains were isolated from 208 of the 342 patients (60.8%) during ICU stay. A comparison of the EK patterns generated from Candida strains isolated from different patients demonstrated a variability of karyotypes, and failed to identify predominant clones colonizing or infecting ICU patients. For 62 of 102 patients colonized with multiple isolates of the same Candida spp., the EK patterns of the Candida strains isolated from individual patients were identical or similar, even when isolated from different anatomical sites, and the patterns remained the same for up to 62 days. A total of 57 episodes of Candida infection occurred in 53 (25.5%) of these 208 patients. Thirty-six episodes (63.1%) of Candida infection were preceded by colonization with the same Candida spp. All infecting strains had identical or similar EK patterns to prior colonizing strains. CONCLUSIONS: While Candida colonization was common in ICU patients, karyotyping did not identify cross-transmission among these patients. Further, only 25.5% of patients with Candida colonization subsequently developed Candida infection. These findings suggest that universal prophylaxis is not warranted in critically ill patients with Candida colonization.

Candida↗