J Mol Biol, 2005 Feb 4, 345(5), 1157 - 69 Epub 2004 Dec 10. Crystal Structure of 2-Enoyl-CoA Hydratase 2 from Human Peroxisomal Multifunctional Enzyme Type 2; Kristian Koski M et al.; 2-Enoyl-CoA hydratase 2 is the middle part of the mammalian peroxisomal multifunctional enzyme type 2 (MFE-2), which is known to be important in the beta-oxidation of very-long-chain and alpha-methyl-branched fatty acids as well as in the synthesis of bile acids . Here, we present the crystal structure of the hydratase 2 from the human MFE-2 to 3A resolution . The three-dimensional structure resembles the recently solved crystal structure of hydratase 2 from the yeast, Candida tropicalis, MFE-2 having a two-domain subunit structure with a C-domain complete hot-dog fold housing the active site, and an N-domain incomplete hot-dog fold housing the cavity for the aliphatic acyl part of the substrate molecule . The ability of human hydratase 2 to utilize such bulky compounds which are not physiological substrates for the fungal ortholog, e.g . CoA esters of C26 fatty acids, pristanic acid and di/trihydroxycholestanoic acids, is explained by a large hydrophobic cavity formed upon the movements of the extremely mobile loops I-III in the N-domain . In the unliganded form of human hydratase 2, however, the loop I blocks the entrance of fatty enoyl-CoAs with chain-length >C8 . Therefore, we expect that upon binding of substrates bulkier than C8, the loop I gives way, contemporaneously causing a secondary effect in the CoA-binding pocket and/or active site required for efficient hydration reaction . This structural feature would explain the inactivity of human hydratase 2 towards short-chain substrates . The solved structure is also used as a tool for analyzing the various inactivating mutations, identified among others in MFE-2-deficient patients . Since hydratase 2 is the last functional unit of mammalian MFE-2 whose structure has been solved, the organization of the functional units in the biologically active full-length enzyme is also discussed. Clin Nephrol, 2004 Dec, 62(6), 473 - 5 Candida tropicalis-associated bilateral renal papillary necrosis and emphysematous pyelonephritis; Wu VC et al.; Although the kidney is often involved in disseminated and localized candidiasis, bilateral emphysematous pyelonephritis (EPN) is infrequently reported . Renal papillary necrosis (RPN) caused by fungi is also rare . We describe a patient with bilateral RPN and EPN caused by Candida tropicalis, who suffered from recurrent hematuria, flank pain, acute fulminant renal failure, and obstruction by a sloughed papilla . He was treated successfully with antifungal therapy and percutaneous nephrostomy (PCN) . This is the first case report of C . tropicalis-associated EPN and RPN. Mycopathologia, 2004 Nov, 158(4), 397 - 405 Epidemiology and molecular typing of Candida isolates from burn patients; Gupta N et al.; This study, spread over a span of 2 years describes Candida infections in burn patients of an Indian hospital . A total of 220 burn patients were monitored and Candida could be isolated from 138 patients . A total of 228 different Candida species were obtained from various body locations of these patients . Species identification revealed that Candida albicans was the most predominant (45) followed by Candida tropicalis(33), Candida glabrata (13.5), C . parapsilosis (4), C . krusei (2.75) and C . kefyr (1.75) . DNA fingerprinting of all C . albicans isolates was done by using CARE-2 probe . Fingerprinting analyses of all the C . albicans strains revealed that strains collected from different patients were different . It is noteworthy that patients with disseminated candidiasis had a similar, but unique strain isolated from all body locations, suggesting a possibility that commensal isolates might be turning pathogenic . Taken together, this is probably the first ever detailed survey of Candidainfections in burn patients in India and is expected to lead to better clinical management of this group of patients. Medicina (B Aires), 2004, 64(2), 152 - 4 {Treatment with caspofungin of Candida tropicalis endocarditis resistant to fluconazol}; del Castillo M et al.; Fungal endocarditis, in particular due to Candida species, requires medical and surgical treatment and amphotericin B is the drug of choice . Caspofungin is an echinocandin very effective against Candida and Aspergillus . We present a patient with Candida tropicalis endocarditis, fluconazol resistant, treated with caspofungin, on a compassional basis as a result of adverse effects with amphotericin B . The patient had a microbiological response. Pediatr Infect Dis J, 2004 Dec, 23(12), 1093 - 7 Caspofungin therapy of neonates with invasive candidiasis; Odio CM et al.; BACKGROUND: Invasive candidiasis is an increasing problem in neonatal intensive care units worldwide and is an important cause of morbidity, mortality and prolongation of hospital stay . Despite administration of amphotericin B, invasive candidiasis in neonates is sometimes complicated by persistent fungemia and refractory invasive candidiasis . The problem has been augmented by the increasing prevalence of non-albicans species that often are resistant to fluconazole and to amphotericin B . POPULATION AND METHODS: The population consisted of 1 term and 9 premature neonates with invasive candidiasis caused by Candida albicans (n = 4), Candida parapsilosis (n = 3), Candida tropicalis (n = 2) and Candida glabrata (n = 1) . Despite initial therapy with deoxycholate amphotericin B, blood cultures remained positive in all patients for 13-49 days . Invasive candidiasis progressed to meningitis and enlarging renal Candida bezoars in the kidney of one patient and an enlarging atrial vegetation in another . Another patient developed severe hypokalemia refractory to potassium supplementation . Two of the C . albicans and all of the non-albicans Candida isolates were resistant to fluconazole; the C . glabrata isolate was resistant to amphotericin B . Amphotericin B was discontinued and caspofungin initiated in all patients in a dosage of 1 mg/kg/d for 2 days followed by 2 mg/kg/d . RESULTS: All positive blood cultures cleared between 3 and 7 days after initiation of caspofungin, the atrial vegetation resolved and the renal Candida bezoars disappeared . Renal and hepatic function tests did not show any values above normal throughout caspofungin therapy . There were no attributable clinical adverse events during the administration of caspofungin in any of the patients . CONCLUSIONS: Caspofungin was effective, safe and well-tolerated as an alternative therapy for persistent and progressive candidiasis in those neonates who were unresponsive to or intolerant of deoxycholate amphotericin B. Rev Esp Quimioter, 2004 Sep, 17(3), 257 - 62 {Study of in vitro activity of caspofungin on non-Candida albicans yeast strains determined by two methods: M27-A2 and EUCAST}; Romero M et al.; The in vitro activity of caspofungin against 147 non-Candida albicans yeasts isolated from blood culture was studied using two broth microdilution methods: M27-A2 and EUCAST . The minimum concentrations that produced a growth inhibition of > or = 50% and of 100% (MIC2 and MIC0, respectively) and a reduction in the number of viable colonies > or = 99% versus the initial inoculum (MFC) were determined for all strains . Caspofungin demonstrated good activity (MIC2 and MIC0 < or = 2 mg/l for 90% of the strains) against the species studied, including those that are normally resistant or have a high percentage of azole resistance (Candida krusei, Candida glabrata, Candida tropicalis) . Nevertheless, the antifungal activity is lower (MFC < or = 2 mg/l in 73.47% of the strains with the M27-A2 method versus 62.59% with EUCAST), particularly for Candida guilliermondii and Candida parapsilosis . The two methods tested demonstrated good correlation for the MIC and lower correlation for the MFC . Essential agreement and concordance (+/- 2 log) between both methods for all strains tested were: 73.47% and 93.20% for MIC2; 74.8% and 91.84% for MIC0; and 57.1% and 74.15% for MFC, respectively . MICs determined by the EUCAST method are one- to three-fold dilutions lower, while the MFC are higher than those obtained by the M27-A2 method. J Dermatol Sci, 2005 Jan, 37(1), 21 - 8 Epub 2004 Dec 09. Epidemiological study of Candida species in cutaneous candidiasis based on PCR using a primer mix specific for the DNA topoisomerase II gene; Kamiya A et al.; BACKGROUND:: We have previously reported a PCR-based identification system for pathogenic fungi by targeting the DNA topoisomerase II gene, in which primer mixes specific for this gene were used for the PCR amplifications . OBJECTIVE:: To test the potential of the PCR using primer mix that is specific for the DNA topoisomerase II gene and are designated as PsVIc, for rapid identification of Candida species involved in cutaneous candidiasis, and to define the relation between Candida species and the infection lesion . METHODS:: Scales from 48 patients with cutaneous candidiasis were cultured on GYEP agar plates, and the genomic DNAs were purified from the colonies and used as DNA templates for PCR amplifications . Candida was identified as individual species based on the sizes of the PCR products generated in the PCR amplifications using PsVlc . RESULTS:: Four Candida species (five genotypes; Candida albicans, Candida glabrata, Candida parapsilosis I, Candida parapsilosis II and Candida tropicalis II) were identified in the patients' scales . In 19 of the patients (39.6%), multiple PCR products (two or three bands) were amplified in a DNA sample, especially derived from scales at the groin of bed-ridden older patients using napkins . CONCLUSION:: The PCR-based identification using the primer mix was useful for an epidemiological study of Candida species in cutaneous candidiasis. J Antimicrob Chemother . 2004 Dec 23; {Epub ahead of print} In vitro susceptibilities of bloodstream isolates of Candida species to six antifungal agents: results from a population-based active surveillance programme, Barcelona, Spain, 2002-2003; Cuenca-Estrella M et al.; The antifungal drug susceptibilities of 351 isolates of Candida species, obtained through active laboratory-based surveillance in the period January 2002-December 2003, were determined (Candida albicans 51%, Candida parapsilosis 23%, Candida tropicalis 10%, Candida glabrata 9%, Candida krusei 4%) . The MICs of amphotericin B, flucytosine, fluconazole, itraconazole, voriconazole and caspofungin were established by means of the broth microdilution reference procedure of the European Committee on Antibiotic Susceptibility Testing . Amphotericin B and flucytosine were active in vitro against all strains . A total of 24 isolates (6.8%) showed decreased susceptibility to fluconazole (MIC >/= 16 mg/L) and 43 (12.3%) showed decreased susceptibility to itraconazole (MIC >/= 0.25 mg/L) . Voriconazole and caspofungin were active in vitro against the majority of isolates, even those that were resistant to fluconazole. Antimicrob Agents Chemother, 2005 Jan, 49(1), 52 - 6 Antifungal activities of R-135853, a sordarin derivative, in experimental candidiasis in mice; Kamai Y et al.; The activities of R-135853, a novel sordarin derivative that possesses a 1,4-oxazepane ring moiety, were evaluated in vitro and in vivo . R-135853 exhibited potent in vitro activities against Candida albicans (fluconazole-susceptible strains), Candida glabrata, Candida tropicalis, and Cryptococcus neoformans, with MICs at which 90% of isolates were inhibited of 0.03, 1, 0.5, and 0.5 microg/ml, respectively . R-135853 also exhibited potent activities against fluconazole-susceptible dose-dependent and fluconazole-resistant strains of C . albicans, with MICs ranging from 0.03 to 0.06 mug/ml . However, R-135853 exhibited weak or no activity against Candida parapsilosis, Candida krusei, and Aspergillus spp . R-135853 exhibited dose-dependent efficacy against experimental murine hematogenous candidiasis induced by C . albicans when it was administered by both the subcutaneous and the oral routes and reduced viable cell counts in the kidneys significantly when it was administered at 50 mg/kg of body weight/dose (administration three times a day) . In this model, R-135853 also exhibited dose-dependent efficacy by single oral administration . Subcutaneous administration of R-135853 exhibited dose-dependent efficacy against experimental murine esophageal candidiasis induced by fluconazole-resistant C . albicans, against which fluconazole at 50 mg/kg/dose was ineffective, and reduced viable cell counts in the esophagus significantly when it was administered at 10 and 50 mg/kg/dose . R-135853 eradicated C . albicans from the esophagi of one and four of five mice when it was administered at 10 and 50 mg/kg/dose, respectively . These results suggest that R-135853 is promising for the treatment of disseminated or mucosal candidiasis, including fluconazole-refractory infections. Am Fam Physician, 2004 Dec 1, 70(11), 2125 - 32 Management of vaginitis; Owen MK et al.; Common infectious forms of vaginitis include bacterial vaginosis, vulvovaginal candidiasis, and trichomoniasis . Vaginitis also can occur because of atrophic changes . Bacterial vaginosis is caused by proliferation of Gardnerella vaginalis, Mycoplasma hominis, and anaerobes . The diagnosis is based primarily on the Amsel criteria (milky discharge, pH greater than 4.5, positive whiff test, clue cells in a wet-mount preparation) . The standard treatment is oral metronidazole in a dosage of 500 mg twice daily for seven days . Vulvovaginal candidiasis can be difficult to diagnose because characteristic signs and symptoms (thick, white discharge, dysuria, vulvovaginal pruritus and swelling) are not specific for the infection . Diagnosis should rely on microscopic examination of a sample from the lateral vaginal wall (10 to 20 percent potassium hydroxide preparation) . Cultures are helpful in women with recurrent or complicated vulvovaginal candidiasis, because species other than Candida albicans (e.g., Candida glabrata, Candida tropicalis) may be present . Topical azole and oral fluconazole are equally efficacious in the management of uncomplicated vulvovaginal candidiasis, but a more extensive regimen may be required for complicated infections . Trichomoniasis may cause a foul-smelling, frothy discharge and, in most affected women, vaginal inflammatory changes . Culture and DNA probe testing are useful in diagnosing the infection; examinations of wet-mount preparations have a high false-negative rate . The standard treatment for trichomoniasis is a single 2-g oral dose of metronidazole . Atrophic vaginitis results from estrogen deficiency . Treatment with topical estrogen is effective. Eur J Clin Microbiol Infect Dis, 2004 Oct, 23(10), 745 - 50 Neonatal candidiasis: analysis of epidemiology, drug susceptibility, and molecular typing of causative isolates; Roilides E et al.; A prospective observational study of invasive candidiasis was conducted in the neonatal intensive care unit of Aristotle University in Hippokration Hospital between 1994 and 2000 . During this period, 59 neonates developed invasive candidiasis (58 cases of candidemia and 1 case of peritonitis), resulting in an overall incidence of 1.28% that showed a decreasing trend over the study period . Eleven (18.6%) cases developed within the first week of life and the others within a mean (+/-SEM) of 13.4+/-1.7 days after birth . The three most frequent causative species were Candida albicans (65.5%), Candida parapsilosis (15.5%), and Candida tropicalis (7%) . C . albicans was the predominant species between 1994 and 1998, whereas, non-albicans Candida spp., particularly C . parapsilosis, were the most frequent species during the period 1999-2000 (P<0.001) . While the overall mortality due to candidemia was 29% (17 of 59 cases), mortality associated with C . albicans and C . parapsilosis was 39.5% and 11.1%, respectively (P=0.032), and that observed in the 1999-2000 period was 0% (P=0.011) . Virtually all isolates were susceptible to amphotericin B, flucytosine, fluconazole, and itraconazole, and no increases in minimal inhibitory concentrations were observed during these years . With the exception of a limited cluster of cases due to genotypically identical isolates, no clonal relation of C . albicans isolates was found . Moreover, no clonal persistence of C . albicans and no decrease in antifungal drug susceptibility occurred over the 6-year study period . Non-albicans Candida spp., mostly C . parapsilosis, have emerged as important pathogens in neonatal intensive care units, with infected patients having better outcomes as compared to patients infected with C . albicans. J Microbiol Immunol Infect, 2004 Dec, 37(6), 335 - 42 In vitro antifungal susceptibility testing of Candida blood isolates and evaluation of the E-test method; Lu JJ et al.; Fungal infections have dramatically increased in recent years, along with the increase of drug-resistant isolates in immunocompromised patients . Ninety eight Candida species obtained from blood cultures at the Tri-Service General Hospital, Taiwan, from 1998 to 2000 were studied . These included 50 Candida albicans, 13 Candida glabrata, 24 Candida tropicalis and 11 Candida parapsilosis isolates . To investigate their susceptibility to commonly used antifungal drugs, minimum inhibitory concentrations (MIC) of amphotericin B, fluconazole, flucytosine, and ketoconazole were determined . Both the National Committee for Clinical Laboratory Standards reference broth macrodilution method and E-test were used in parallel . Ninety five isolates (95/98, 96.94%) were susceptible to amphotericin B at a concentration < or = 1 microg/mL . All isolates (100%, 98/98) were susceptible to flucytosine . Approximately 30% of these Candida isolates were resistant to fluconazole . The MIC for 90% of isolates (MIC90) values for both methods for these isolates were 0.5 microg/mL for amphotericin B, 32 microg/mL for fluconazole, 0.25 microg/mL for flucytosine (0.125 microg/mL by E-test method), and 4 microg/mL for ketoconazole . MIC for 50% of isolates (MIC50) values for these agents were 0.25, 2, 0.06, and 0.06 microg/mL, respectively . The essential agreement of MIC values within 2 dilutions for the 2 methods was 99.0% for amphotericin B, 90.8% for ketoconazole, 92.9% for fluconazole, and 91.8% for flucytosine . This study showed that E-test has equivalent performance to the broth macrodilution method and can be used as an alternative MIC technique for antifungal susceptibility testing. Arch Oral Biol, 2005 Jan, 50(1), 33 - 37 Prevalence of Candida species in Turkish children: relationship between dietary intake and carriage; Kadir T et al.; In this study, the prevalence and intensity of Candida species were evaluated in 300 healthy Turkish children aged between 0 and 12 years . The candidal carriage in 26 children who were fed only with breast milk and 38 children who were fed with both breast milk and bottle milk or other fluids was also examined . Oral samples cultured for fungal growth and Candida species were identified using germ tube test, chlamydospore formation test and API 20C AUX system . The results demonstrated that the prevalence of oral candidal carriage in 300 healthy children was 26.3% . Candida albicans was the most frequently isolated yeast (84.8% of the isolates) . The other yeasts were identified as Candida parapsilosis, Candida krusei, Candida kefyr, Candida famata, and Candida tropicalis . It was also observed that the frequency of carriage varied as a function of age . The prevalence of carriage in children who were fed with both breast milk and bottle milk or other fluids was 18.5%, while in children fed only with breast milk was 0% . This finding supports previously reported observations that there may be intrinsic differences in oral carriage of Candida species between different ages and populations and type of dietary intake may affect frequency of carriage. Biosens Bioelectron, 2005 Jan 15, 20(7), 1263 - 9 An amperometric microbial biosensor development based on Candida tropicalis yeast cells for sensitive determination of ethanol; Akyilmaz E et al.; Different branchs of industry need to use ethanol in their production and some progress and not only the industry also to determine ethanol sensitively, accurately, fast and economical is very important . For the sensitive determination of ethanol a new amperometric biosensor based on Candida tropicalis cells, which contains alcohol oxidase enzyme, immobilized in gelatin by using glutaraldehyde was developed . In the study, before the microbial biosensor construction C . tropicalis cells were activated and cultured in a culture medium . By using gelatine and glutaraldehyde (0.1%) C . tropicalis cells obtained in logarithmic phase were immobilized and fixed on a pretreated teflon membrane of a dissolved oxygen probe . Ethanol determination is based on the assay of the differences on the respiration activity of the cells on the oxygenmeter in the absence and the presence of ethanol . The microbial biosensor response was depend linearly on ethanol concentration between 0.5 and 7.5mM with 2min response time . In the optimization studies of the microbial biosensor the most suitable microorganism amount were found as 4.42mgcm(-2) and also phosphate buffer (pH:7.5; 50mM) and 30 degrees C were obtained as the optimum working conditions . In the characterization studies of the microbial biosensor some parameters such as substrate specificity, interference effects of some substances on the biosensor response, operational and storage stability were carried out. J Antimicrob Chemother, 2005 Jan, 55(1), 110 - 4 Epub 2004 Dec 01. In vitro activity of bergamot natural essence and furocoumarin-free and distilled extracts, and their associations with boric acid, against clinical yeast isolates; Romano L et al.; OBJECTIVES: There is very little information, to date, on the antifungal activity of bergamot oil . In this study, we investigated the in vitro activity of three bergamot oils (natural essence, furocoumarin-free extract and distilled extract) against clinically relevant Candida species . We studied the two derivatives, components of Italian pharmaceutical products, that are supposed to be less toxic than the essential oil . METHODS: In vitro susceptibility of 40 clinical isolates of Candida spp . (Candida albicans, n=20; Candida glabrata, n=13; Candida krusei, n=4; Candida tropicalis, n=2; Candida parapsilosis, n=1), associated with symptomatic and asymptomatic vulvovaginal candidiasis, was determined using a modification of the NCCLS M27-A2 broth microdilution method . MICs were evaluated for each of the oils alone and combined with sub-inhibitory concentrations of the well-known antiseptic, boric acid . To boric acid, all isolates had MIC values ranging from 0.094% to 0.187% (w/v) . RESULTS: At 24 h readings, the MIC(90 )s (for all isolates) were (v/v): 5% for natural essence of bergamot, 2.5% for the furocoumarin-free extract, and 1.25% for the distilled extract . At the 48 h reading, these values increased to >10%, 5% and 2.5%, respectively . At both readings, MIC(90 )s for all oil+boric acid combinations were significantly lower than corresponding values for the oils alone (P <0.05) . CONCLUSIONS: These data indicate that bergamot oils are active in vitro against Candida spp., suggesting their potential role for the topical treatment of Candida infections. Langmuir, 2004 Dec 7, 20(25), 10949 - 55 Microbial adhesion to poly(ethylene oxide) brushes: influence of polymer chain length and temperature; Roosjen A et al.; Glass surfaces were modified by end-grafting poly(ethylene oxide) (PEO) chains having molecular weights of 526, 2000, or 9800 Da . Characterization using water contact angles, ellipsometry, and X-ray photoelectron spectroscopy confirmed the presence of the PEO brushes on the surface with estimated lengths in water of 2.8-, 7.5-, and 23.7-nm, respectively . Adhesion of two bacterial (Staphylococcus epidermidis and Pseudomonas aeruginosa) and two yeast (Candida albicans and Candida tropicalis) strains to these brushes was studied and compared to their adhesion to bare glass . For the bacterium P . aeruginosa and the yeast C . tropicalis, adhesion to the 2.8-nm brush was comparable to their adhesion on bare glass, whereas adhesion to the 7.5- and 23.7-nm brushes was greatly reduced . For S . epidermidis, adhesion was only slightly higher to the 2.8-nm brush than that to the longer brushes . Adhesion of the yeast C . albicans to the PEO brushes was lower than that to glass, but no differences in adhesion were found between the three brush lengths . After passage of an air bubble, nearly all microorganisms adhering to a brush were removed, irrespective of brush length, whereas retention of the adhering organisms on glass was much higher . No significant differences were found in adhesion nor retention between experiments conducted at 20 and those conducted at 37 degrees C. Yeast, 2004 Dec, 21(16), 1335 - 42 Alcohol-mediated haemolysis in yeast; Shuster A et al.; Although yeast are generally non-haemolytic, we have found that addition of alcohol vapour confers haemolytic properties on many strains of yeast and other fungi . We have called this phenomenon 'microbial alcohol-conferred haemolysis' (MACH) . MACH is species- and strain-specific: whereas all six Candida tropicalis strains tested were haemolytic in the presence of ethanol, none among 10 C . glabrata strains tested exhibited this phenomenon . Among 27 C . albicans strains and 11 Saccharomyces cerevisiae strains tested, ethanol-mediated haemolysis was observed in 11 and 4 strains, respectively . Haemolysis is also dependent on the alcohol moiety: n-butanol and n-pentanol could also confer haemolysis, whereas methanol and 2-propanol did not . Haemolysis was found to be dependent on initial oxidation of the alcohol . Reduced haemolysis was observed in specific alcohol dehydrogenase mutants of both Aspergillus nidulans and S . cerevisiae . MACH was not observed during anaerobic growth, and was reduced in the presence of pararosaniline, an aldehyde scavenger . Results suggest that initial oxidation of the alcohol to the corresponding aldehyde is an essential step in the observed phenomenon. J Antimicrob Chemother, 2004 Dec, 54(6), 1051 - 6 Epub 2004 Nov 25. Initial results from a longitudinal international surveillance programme for anidulafungin (2003); Messer SA et al.; OBJECTIVES: This longitudinal study evaluated the in vitro activity of anidulafungin against 880 clinical yeast isolates and 68 mould isolates from 64 medical centres in North America, Latin America and Europe . METHODS: MICs of anidulafungin, amphotericin B, 5-fluorocytosine, fluconazole, itraconazole, ketoconazole and voriconazole were determined using reference method (M27-A2) guidelines . The M38-A reference method was used for the filamentous fungi, including determination of minimum effective concentrations (MECs) of anidulafungin . RESULTS: Anidulafungin was more active when compared with fluconazole and itraconazole for Candida albicans (MIC(90), 0.06 mg/L), Candida tropicalis (MIC(90), 0.06 mg/L), Candida glabrata (MIC(90), 0.12 mg/L), Candida krusei (MIC(90), 0.06 mg/L) and Candida lusitaniae (MIC(90), 1 mg/L) as well as the less-often encountered yeast species . Anidulafungin was less active against Candida parapsilosis, Candida guilliermondii and Candida famata (MIC(50), 1-2 mg/L) . Anidulafungin also exhibited excellent activity against all Aspergillus spp . (MEC(90), </=0.03 mg/L) . Anidulafungin was also evaluated comparing two end point reading criteria and two incubation intervals . Data indicate that longer incubation periods do not significantly influence overall MIC ranges . These international surveillance results for anidulafungin confirm the activity observed in studies of smaller numbers of isolates. Prikl Biokhim Mikrobiol, 2004 Sep-Oct, 40(5), 533 - 5 {Search for yeast producers of brassylic and sebacic fatty acids}; Mild detergent treatment of Candida tropicalis reveals a NADPH-dependent reductase in the crude membrane fraction et al.; Department of Applied Microbiology, Lund University, PO Box 124, 221 00 Lund, SwedenThis study demonstrated the occurrence of a NADPH-dependent exo-alcohol reductase in the crude membrane fraction of Candida tropicalis . Cytosolic endo-alcohol reductase activity could be separated from the membrane-bound exo-alcohol activity by means of detergent treatment, enabling the preparation of pure exo-alcohol via the enzymatic conversion of the bicyclic diketone, bicyclo{2.2.2}octane-2,6-dione . The exo-alcohol reductase is, to our knowledge, the first membrane-bound NADPH-dependent reductase accepting a xenobiotic carbonyl substrate that was not a steroid . When C . tropicalis was grown on D-sorbitol, a two-fold increase in the exo-reductase activity was observed as compared to when grown on glucose . An in silico comparison at the protein level between putative xenobiotic carbonyl reductases in Candida albicans, C . tropicalis and Saccharomyces cerevisiae was performed to explain why Candida species are often encountered when screening yeasts for novel stereoselective reduction properties . C . albicans contained more reductases with the potential to reduce xenobiotic carbonyl compounds than did S . cerevisiae . C . tropicalis had many membrane-bound reductases (predicted with the bioinformatics program, TMHMM), some of which had no counterpart in the two other organisms . The exo-reductase is suspected to be either a beta-hydroxysteroid dehydrogenase or a polyol dehydrogenase from either the short chain dehydrogenase family or the dihydroflavonol reductase family. Diagn Microbiol Infect Dis, 2004 Nov, 50(3), 179 - 85 Tolerance to amphotericin B in clinical isolates of Candida tropicalis; Barchiesi F et al.; A broth microdilution method was used for testing amphotericin B against 33 clinical isolates of Candida tropicalis . All isolates were in vitro susceptible to the polyene (MIC {minimal inhibitory concentration} < or = 1.0 microg/mL) . However, when the isolates were cultured in a medium containing amphotericin B at a concentration of 1.5 microg/mL, a wide interstrain variation of growth rate was observed . Five isolates (15%) proved to be highly tolerant to the drug and grew at a frequency ranging from 1 x 10(-1) to 2 x 10(-2) . Twenty-three isolates (70%) grew at a frequency ranging from 1 x 10(-5) to 1 x 10(-8) . The remaining five isolates (15%) failed to grow in drug-containing medium . In general, this growth variation was not associated with amphotericin B MICs displayed by the single isolates . In addition, the strains grown in drug-containing medium did not represent amphotericin B-resistant mutants, as shown by the maintenance of MICs similar to those of their respective parent isolates . Killing experiments conducted in selected isolates confirmed a variation of fungicidal activity of amphotericin B . To see whether this phenomenon was associated with a variation of amphotericin B response in vivo, we established an experimental model of systemic murine candidiasis in CD1 mice by intravenous injection of cells belonging to Candida tropicalis 3147 (growth rate at a frequency of 1 x 10(-1) in amphotericin B medium) and Candida tropicalis 4055 (no growth) . Low (0.3 mg/kg/day) and high (1 mg/kg/day) doses of amphotericin B were both effective at reducing the fungal burdens in the kidneys of mice infected with either strain (p, 0.01 to 0.02) . However, whereas the burden of mice infected with isolate 3147 and treated with the polyene at 0.3 mg/kg/day was reduced by 1.2 +/- 0.25 (mean +/- standard deviation) log10 cfu/g compared to untreated mice, the same dosing regimen yielded a burden reduction of 2.6 +/- 0.07 log10 cfu/g in mice infected with isolate 4055 (p < 0.001) . Similarly, amphotericin B at 1 mg/kg/day yielded a burden reduction of 1.8 +/- 0.20 vs . 2.5 +/- 0.30 log10 cfu/g in mice infected with isolates 3147 and 4055, respectively (p < 0.001) . Our data revealed a variable pattern of tolerance to amphotericin B among isolates of Candida tropicalis and showed that this phenomenon might influence the rate of organ clearance during therapy. Rev Iberoam Micol, 2004 Jun, 21(2), 82 - 4 {Rapid identification of Candida glabrata using a new commercial kit}; Peman J et al.; Candida glabrata is an emergent pathogen with diminished susceptibility to azoles, thus a rapid identification of this yeast could be of help to choose the appropriate treatment . GLABRATA RTT (Fumouze Diagnostics, France) is a new C . glabrata identification test . To evaluate its utility in the clinical laboratory daily routine, we prospectively tested 168 yeasts isolated in our hospital . GLABRATA RTT results had a sensitivity of 98.4% and a specificity of 100% . The combination of CHROMagar Candida isolation medium and GLABRATA RTT test allowed the identification of the four most common species in the clinical practice (Candida albicans, Candida tropicalis, Candida krusei and C . glabrata). Rev Iberoam Micol, 2004 Jun, 21(2), 79 - 81 Posaconazole therapy for severe abdominal candidiasis: a case report; Tobon AM et al.; We report the successful treatment of a fluconazole-resistant intra-abdominal Candida infection (Candida albicans and Candida tropicalis) with posaconazole (SCH56592) in a 68-year-old woman with a recent history of intra-abdominal surgery. Rev Iberoam Micol, 2004 Jun, 21(2), 63 - 9 In-vitro activity of 5-fluorocytosine against 1,021 Spanish clinical isolates of Candida and other medically important yeasts; Quindos G et al.; 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. Appl Biochem Biotechnol, 2004 Oct, 119(1), 13 - 30 Effect of detoxification of dilute-acid corn fiber hydrolysate on xylitol production; Buhner J et al.; Four different detoxification methods were evaluated for the production of xylitol from corn fiber dilute-acid hydrolysate using Candida tropicalis . Although C . tropicalis could ferment the dilute partially neutralized hydrolysate to xylitol in low yields (0.1 g/g), it could not ferment the concentrated hydrolysate . Overliming, calcium hydroxide neutralization, neutralization combined with activated charcoal, and overliming combined with activated charcoal methods were used to improve the fermentation of the concentrated hydrolysates . The partial neutralization combined with activated charcoal treatment was the most effective method with respect to xylitol yield and productivity . The highest xylitol yield (0.4 g of xylitol/g of xylose) was obtained for the highest concentration of hydrolysate (three times the original) that had been treated with calcium hydroxide and activated charcoal . The corresponding productivity was 0.23 g/(L x h) . Overliming caused reduction in xylitol yield. Rev Iberoam Micol, 2001 Dec, 18(4), 197 - 9 {Identification of Candida albicans using the chromogenic medium Albicans ID.}; Godoy P et al.; The correct identification of the microrganism is the base for epidemiological studies and treatment of infections . The aim of our study was to evaluate the efficacy of the chromogenic media Albicans ID (bioMerieux, France) in the identification of Candida albicans . A total of 190 yeasts strains were evaluated in the study . A rate of 100% of all C . albicans (80) and Candida dubliniensis (five) strains exhibited blue color . Nevertheless, the blue color was also observed with cultures of Candida rugosa (3/5) and Candida tropicalis (3/17) . Albicans ID cromogenic media presented specificity rate of 90% and positive and negative predictive values of 88% and 100%, respectivly, in the identification of C . albicans. Rev Iberoam Micol, 2001 Dec, 18(4), 171 - 3 Effect of contraceptives on the prevalence of vaginal colonization with Candida species in Edo State, Nigeria; Enweani IB et al.; High vaginal swabs (HVS) obtained from 500 volunteers in Edo State, Nigeria which comprised 394 contraceptive users and 106 non-contraceptive users were screened for the prevalence of Candida species using standard procedures . Results revealed the isolation of Candida species in 246 of volunteers . These included Candida albicans 174 (38.4%), Candida pseudotropicalis 20 (4%), Candida stellatoidea 15 (3%), Candida krusei nine (1.8%), Candida guilliermondii 12 (2.4%), Candida tropicalis 11 (2.2%) and Candida glabrata five (1%) . Of the 394 contraceptive users, 203 (51.5%) had Candida species isolated from them compared to 43 (40.6%) from 106 non-contraceptive users . There was significant relationship (P<0.001) between the type of contraceptive used and the prevalence of vaginal colonization . Age and marital status of the volunteers sampled had significant relationship (P<0.001) with the prevalence of vaginal colonization . Results have revealed an association between use of contraceptive and the prevalence of vaginal colonization in our environment. J Environ Sci (China), 2004, 16(4), 690 - 3 Straw bio-degradation by acidogenic bacteria and composite fungi; Zhang KQ et al.; A composite microbial system, including a strain of Candida tropicalis (W3), a strain of Lactobacillus plantarm(WY3) and three strains of basidiomycete pL104, pL113 and C33, was chosen to degrade corn straw . The final pH was acid owing to the inoculation of acidogenic bacteria, and under this condition the composite fungi system could produce complex enzyme to destroy the compact structure of corn straw . The experimental results showed that the biomass of composite fungi could reach up to maximum when the pH value was 4.5 . Through the bio-degradation by combining acidogenic bacteria with the composite fungi system, the cellulose, hemi-cellulose and lignin degradation rates of corn straw powder were 26.36%, 43.30% and 26.96%, respectively . And the gross crude protein content increased 60.41% . This study provided the evidence for the feasibility of developing a composite microbial system with high capability of degrading straw lignocelluloses in order to make reasonable use of straw resource and protect rural eco-environment. Oral Microbiol Immunol, 2004 Dec, 19(6), 347 - 51 Oral Candida isolates in patients undergoing radiotherapy for head and neck cancer: prevalence, azole susceptibility profiles and response to antifungal treatment; Belazi M et al.; Oral pseudomembranous candidiasis and mucositis were assessed in 39 patients receiving a total dose of 39-70 Gy radiotherapy for head and neck cancer . Mucositis was scored using the Radiation Therapy Oncology Group criteria, and oral candidiasis was diagnosed on the basis of clinical evaluation and quantitative laboratory findings . Radiation-induced mucositis was observed in 9/39 patients . Only 3/39 patients discontinued radiotherapy due to acute severe mucosal effects . Candidiasis (colony-forming units 35 to > or = 60/lesion) associated with mucositis was diagnosed in 30/39 patients: the most frequent aetiology of the infection was Candida albicans (n = 23), followed by Candida glabrata (n = 3), Candida krusei (n = 2), Candida tropicalis (n = 1) and Candida kefyr (n = 1) . Patients with confirmed oral pseudomembranous candidiasis were treated with either fluconazole 200 mg/day or itraconazole 200 mg/day for 2 weeks . Clinical improvement and concomitant negative Candida cultures (mycologic cure) were the criteria determining a response to antifungal treatment . Etest revealed very low voriconazole MICs (0.004-0.125 microg/ml) for all isolates, and fluconazole resistance for eight C . albicans strains (MIC > 64 microg/ml) and for the C . krusei isolates (MIC > 32 microg/ml) . The same strains showed itraconazole susceptibility dose dependence (MIC 0.5 microg/ml) . Despite the itraconazole susceptible dose dependent MIC readings, all patients with oral pseudomembranous candidiasis caused by these strains responded to antifungal treatment with 200 mg/day itraconazole . Oral mycologic surveillance of patients undergoing radiotherapy for head and neck malignancies and susceptibility testing of isolates may be indicated in cases with mucositis-associated confirmed oral pseudomembranous candidiasis to ensure prompt administration of targeted antifungal treatment . On the basis of the low MIC values found, clinical evaluation of voriconazole is indicated for management of oral pseudomembranous candidiasis refractory to other azoles. Rev Iberoam Micol, 2001 Jun, 18(2), 60 - 4 Inter and intra-specific genetic variability of oral Candida species; Ribeiro-Rosa EA et al.; In this report, strains of five different Candida species (Candida albicans, Candida guilliermondii, Candida tropicalis, Candida krusei, and Candida parapsilosis) isolated from healthy human oral cavities as well as their respective type-strains were used in order to establish the genetic diversity existing among the different species and within a certain species, by the analysis of their electrophoretic alloenzyme patterns . These profiles were analyzed for their band positions in the gels, which allowed to group the strains of the same species in species-specific clusters and to treat them as conspecific populations . A total of thirteen enzymatic loci were obtained (ACO, ADH1, ADH2, CAT, G6PDH, GDH, GOT, IDH1, IDH2, LAP, LDH, PER, and SOD) . The allelic frequencies (p) and the heterozygosity (h) for all the thirteen loci were determined by diversity index formulas . The GST index is the estimated proportion of genetic diversity that was applied in order to establish inter and intra populational diversity, which, for our results, indicated that 37.75% of total genetic diversity was attributable to differences among the species and the remaining 62.25% was attributable to differences within these populations . An Euclidian distance dendrogram for the different conspecific populations was built, showing that C . guilliermondii grouped first with C . tropicalis and thus formed a expanded cluster with C . albicans . This cluster combined later with another one composed by C . parapsilosis and C . krusei . Comparing our results to the others that were obtained by different molecular techniques, we have observed that the clustering hierarchies follow different paths of organization, varying according to the methodology employed. Rev Iberoam Micol, 2002 Mar, 18(1), 23 - 28 {Evaluation of a new chromogenic medium (Candida IDtrade mark) for the isolation and presumptive identification of Candida albicans and other medically important yeasts.}; Quindos G et al.; Candidiasis is a frequent human infection caused mainly by Candida albicans . However, other species are emerging as important pathogens, as Candida glabrata, Candida parapsilosis, Candida tropicalis, Candida krusei or Candida guilliermondii . Rapid identification of clinical isolates could facilitate diagnosis and treatment . Candida IDtrade mark (bioMerieux, Spain) is a new medium for the isolation and presumptive identification of yeasts: C . albicans grows as blue colonies, and C . tropicalis, C . guilliermondii, Candida kefyr and Candida lusitaniae as pink ones . The utility of Candida ID was evaluated with more than 700 clinical isolates and type culture collection strains from different genera including Candida, Cryptococcus, Saccharomyces, and Rhodotorula . Presumptive identification was confirmed by germ tube test, microscopic morphology and chlamydoconidia production on corn meal agar and carbohydrate assimilation on API-ATB ID 32C or Vitek (bioMerieux) . Growth on Candida ID was rapid (18-24 h) for most of the yeast strains tested . Sensitivity and specificity of identification of C . albicans was significantly high (>98%), since a very low number of isolates were found to be false negative or false positive . A better result was obtained for species growing as pink colonies (>99.5%) . Detection of different species of medical important yeasts was easy with Candida ID, as perfectly distinct colors and textures of colonies were observed on this medium . Candida ID allowed the discrimination between C . glabrata (creamy and smooth) and C . krusei (rough and white) colonies . Other species showed different colony textures and colours, white being the predominant colour . Candida ID was very useful for the presumptive identification C . albicans isolates. Biochem Biophys Res Commun, 2004 Nov 5, 324(1), 25 - 30 Site-directed mutagenesis to enable and improve crystallizability of Candida tropicalis (3R)-hydroxyacyl-CoA dehydrogenase; Ylianttila MS et al.; The N-terminal part of Candida tropicalis MFE-2 (MFE-2(h2Delta)) having two (3R)-hydroxyacyl-CoA dehydrogenases with different substrate specificities has been purified and crystallized as a recombinant protein . The expressed construct was modified so that a stabile, homogeneous protein could be obtained instead of an unstabile wild-type form with a large amount of cleavage products . Cubic crystals with unit cell parameters a=74.895, b=78.340, c=95.445, and alpha=beta=gamma=90 degrees were obtained by using PEG 4000 as a precipitant . The crystals exhibit the space group P2(1)2(1)2(1) and contain one molecule, consisting of two different (3R)-hydroxyacyl-CoA dehydrogenases, in the asymmetric unit . The crystals diffract to a resolution of 2.2A at a conventional X-ray source. Rev Iberoam Micol, 2004 Mar, 21(1), 24 - 8 Phenotypic and genotypic identification of Candida spp . isolated from hospitalized patients; Campos de Pinho Resende J et al.; As candidosis incidence continue to rise, quick laboratory identification of Candida species is becoming increasingly important for a growing population of patients at-risk . RAPD techniques were used on samples of Candida obtained from patients hospitalized at Santa Casa de Misericordia in Belo Horizonte (SCMBH) Brazil, from March 1998 to December 2000 and then compared with the results of phenotypic identification techniques . Two hundred and forty two yeasts were isolated and phenotypically identified as follows: Candida albicans (105), Candida tropicalis (62), Candida parapsilosis (28), Candida glabrata (19), Candida krusei (8), Candida guilliermondii (5) and Candida spp . (15) . Samples from the three most frequent species isolated were selected randomly in order to compare the phenotypic and genotypic analyses . Genotypic analysis using RAPD primer M13 (F/R) displayed the best results of all test samples . There was both agreement and consistency between phenotypic and genotypic analysis using RAPD, demonstrating that is possible to apply this method for the identification of Candida species. Rev Iberoam Micol, 2003 Jun, 20(2), 60 - 3 The distribution frequency of Candida species in the genitourinary tract among symptomatic individuals in Nigerian cities; Okungbowa FI et al.; A clinical survey was carried out in seven cities in the southern part of Nigeria to determine the relative distribution of genitourinary Candida species in symptomatic patients reporting for diagnosis and treatment . Seven Candida species were identified using the CHROMagar Candida method and the API 20C System . Candida species were represented by Candida glabrata (33.7%), Candida albicans (20.1%), Candida tropicalis (18%), Candida guilliermondii (17.8%) Candida pseudotropicalis (5%), Candida parapsilosis (5%), and C . albicans var.stellatoidea (1.2%) . The distribution of these species among the various age groups (15-20, 21-25, 26-30, 31-35, 36-40 and 41 plus years) was statistically insignificant . Out of the 517 positive samples, 182 (35%) were found in the age group 26-30 years, while age 41 plus had the lowest frequency (1.2%) . The results presented show that C . albicans, usually reported to be the most frequently isolated species, is not the main species in the cities studied . With C . glabrata in preponderance, the finding supports recent studies reporting that several pathogenic non-C . albicans species are now being frequently isolated . The level of social activities, such as drug abuse and sexual promiscuity, may be important in the distribution frequency of Candida species in different age groups and locations. J Microbiol, 2004 Jun, 42(2), 80 - 6 Molecular investigation of two consecutive nosocomial clusters of Candida tropicalis candiduria using pulsed-field gel electrophoresis; Rho J et al.; Pulsed-field gel electrophoresis (PFGE) typing was applied to the epidemiological investigation of 21 Candida tropicalis isolates collected from urine specimens of 11 patients and one healthcare worker, in an intensive care unit (ICU) over a 4-month period . Seventeen epidemiologically unrelated strains from 14 patients were also tested to determine the discriminatory power of PFGE . PFGE typing consisted of electrophoretic karyotyping (EK) and restriction endonuclease analysis of genomic DNA (REAG), using two restriction enzymes (BssHII and SfiI) . The EK pattern was the same in all 38 isolates, while REAG using SfiI separated the isolates into nine types . However, 16 different PFGE types were identified by REAG with BssHII, and the same results were obtained when the results of both REAG tests were combined . In serial urinary isolates from 10 patients, all strains from each patient had the same PFGE pattern . While the epidemiologically unrelated strains from 14 patients consisted of 13 different PFGE types, the 20 isolates from the 11 ICU patients fell into only two PFGE types (types C1 and C2), and these apparently originated from the two different outbreaks . All strains of type C1 (n = 12) were isolated from six patients, between November 1999 and January 2000, and all of the type C2 strains (n=8) were isolated from five patients, during January and February 2000 . This study shows two consecutive clusters of C . tropicalis candiduria in an ICU, defined by PFGE typing, and also demonstrates that a PFGE typing method using BssHII is perhaps the most useful method for investigating C . tropicalis candiduria. Infect Control Hosp Epidemiol, 2004 Aug, 25(8), 634 - 40 Genetic relatedness among Candida tropicalis isolates from sporadic cases of fungemia in two university hospitals in Korea; Shin JH et al.; OBJECTIVE: To compare the epidemiology and genetic relatedness of Candida tropicalis isolates causing bloodstream infection (BSI) in two hospitals . SETTING: Two tertiary-care hospitals in Korea . METHODS: A retrospective molecular epidemiologic analysis using pulsed-field gel electrophoresis (PFGE) was performed with 49 C . tropicalis isolates from sporadic cases of BSI . The isolates were collected from 27 patients at Chonnam National University Hospital (CUH) during a 6-year period and 22 patients at Asan Medical Center (AMC) during a 2-year period . RESULTS: Based on the PFGE patterns, the average similarity value (S AB) for the 27 isolates from CUH was 0.84 +/- 0.08, which was significantly higher than that for the 22 isolates from AMC (0.78 +/- 0.06; P < .001) . Of the 49 strains from patients at the 2 hospitals, 9 isolates were placed into 3 subtypes with S AB values of 1.0, which indicated that they were identical . All 9 of these strains were isolated from CUH patients, and each type strain was isolated sporadically during a period ranging from 4 months to 3 years . On comparison of the clinical characteristics of the patients of the 2 hospitals, the CUH strains were isolated more frequently from non-neutropenic patients and patients with central venous catheter-related fungemia; cases from CUH had a better outcome than those from AMC (P < .05) . CONCLUSIONS: These data show that the clinical and epidemiologic characteristics of C . tropicalis fungemia may differ markedly among hospitals and that some cases of C . tropicalis fungemia may be caused by endemic strains within a hospital. South Med J, 2004 Aug, 97(8), 788 - 90 Disseminated Candida tropicalis in a patient with chronic mucocutaneous candidiasis; Dixon TC et al.; Chronic mucocutaneous candidiasis is a heterogeneous group of immunodeficiencies associated with persistent candidal infections . Patients with chronic mucocutaneous candidiasis are rarely associated with systemic infections caused by other fungi, but almost never by Candida . The authors report a case of a 16-year-old with chronic mucocutaneous candidiasis who developed a fungemia with Candida tropicalis. J Mater Sci Mater Med, 2001 May, 12(5), 399 - 405 Conditioning film and environmental effects on the adherence of Candida spp . to silicone and poly(vinylchloride) biomaterials; Jones DS et al.; The reported incidence of colonization of oropharyngeal medical devices with Candida spp . has increased in recent years, although few studies that have systematically examined the adherence of yeast cells to such biomaterials, the primary step in the process of colonization . This study, therefore, examined the effects of oropharyngeal atmospheric conditions (5% v/v carbon dioxide) and the presence of a salivary conditioning film on both the surface properties and adherence of Candida albicans, Candida krusei and Candida tropicalis to PVC and silicone . Furthermore, the effects of the salivary conditioning film on the surface properties of these biomaterials are reported . Growth of the three Candida spp . in an atmosphere containing 5% v/v \hbox{CO}_{2} significantly increased their cell surface hydrophobicity and reduced the zeta potential of C . albicans and C . krusei yet increased the zeta potential of C . tropicalis (p<0.05) . Furthermore, growth in 5% v/v \hbox{CO}_{2} decreased the adherence of C . tropicalis and C . albicans to both PVC and silicone, however, increased adherence of C . krusei (p<0.05) . Pre-treatment of the microorganisms with pooled human saliva significantly decreased their cell surface hydrophobicity and increased their adherence to either biomaterial in comparison to yeast cells that had been pre-treated with PBS (p<0.05) . Saliva treatment of the microorganisms had no consistent effect on microbial zeta potential . Interestingly, adherence of the three, saliva-treated Candida spp . to saliva-treated silicone and PVC was significantly lower than whenever the microorganisms and biomaterials had been treated with PBS (p<0.05) . Treatment of silicone and PVC with saliva significantly altered the surface properties, notably reducing both the advancing and receding contact angles and, additionally, the microrugosity . These effects may contribute to the decreased adherence of saliva-treated microorganisms to these biomaterials . In conclusion, this study has demonstrated the effects of physiological conditions within the oral cavity on the adherence of selected Candida spp . to biomaterials employed as oropharyngeal medical devices . In particular, this study has ominously shown that these materials act as substrates for yeast colonization, highlighting the need for advancements in biomaterial design . Furthermore, it is important that physiological conditions should be employed whenever biocompatibility of oropharyngeal biomaterials is under investigation . J Microbiol Immunol Infect, 2004 Aug, 37(4), 236 - 41 Species distribution and fluconazole susceptibility of Candida clinical isolates in a medical center in 2002; Wang JL et al.; Fluconazole disk-diffusion susceptibility was evaluated in 230 blood isolates and 344 non-blood clinical isolates of Candida spp . collected in 2002 at National Taiwan University Hospital . Up to 93.5% of blood isolates were susceptible to fluconazole, 3% were susceptible dose-dependent, and 3.5% were resistant . The minimum inhibitory concentrations at which 50% of tested isolates were inhibited (MIC50) of fluconazole against Candida blood isolates were highest for Candida glabrata (5 microg/mL), followed by Candida tropicalis (2.4 microg/mL), Candida albicans (2.4 microg/mL), and Candida parapsilosis (0.41 microg/mL) . C . glabrata had less fluconazole-susceptible strains (76.7%) than C . albicans (98.2%), C . tropicalis (98%) and C . parapsilosis (93.8%) {p<0.05} . The proportions of fluconazole resistance in the non-blood isolates of C . albicans, C . glabrata and C . parapsilosis were similar to those of the blood isolates . However, the proportions of fluconazole resistance in the non-blood isolates of C . tropicalis surpassed those of the blood isolates (14.7% vs 2%, p<0.05) . Comparison of species distribution of Candida blood isolates obtained in 2002 to those in 1981-2000 demonstrated that C . albicans remained the leading pathogen, and the proportion of C . albicans in blood isolates was lowest in 1996 (38%) and did not change significantly thereafter . However, the proportion of C . tropicalis increased from 14% during 1981-1993 to 22-23% during 1996-2002 . Overall, the MIC50, MIC90 and the proportion of Candida blood isolates with fluconazole resistance remained stable during 1994-2002. Int J Antimicrob Agents, 2004 Sep, 24(3), 294 - 6 In vitro activity of voriconazole against Candida species isolated in Taiwan; Yang YL et al.; The activity of voriconazole was determined against 285 Candida species consisting of 53 resistant isolates, 43 susceptible-dose dependent and 189 isolates susceptible to fluconazole . The MIC(50) and MIC(90) to fluconazole were 8 and 64 mg/l, respectively . The range of minimum inhibitory concentrations (MICs) to voriconazole was from 0.0325 to 2 mg/l and the MIC(50) and MIC(90) were 0.125 and 0.5 mg/l, respectively . Only 3 of 285 tested isolates had MICs to voriconazole equal to 2 mg/l . A total of 38 isolates, consisted of 3 Candida albicans, 5 Candida krusei, 7 Candida tropicalis and 21 Candida glabrata, had >/= 0.5 mg/l to voriconazole . There was correlation between the susceptibility to fluconazole and voriconazole. J Eur Acad Dermatol Venereol, 2004 Sep, 18(5), 617 - 8 Onychomycosis in a premature infant caused by Candida tropicalis; Chun DK et al.; We report a case of onychomycosis caused by Candida tropicalis in a 107-day-old infant . Res Microbiol, 2004 Sep, 155(7), 579 - 86 In vitro activity of essential oil from Ocimum gratissimum L . against four Candida species; Nakamura CV et al.; Development of effective strategies for treatment of candidiasis and other fungal diseases has posed a challenge, considering the increase in opportunistic fungal infections in HIV-positive and immunocompromised patients . The in vitro antifungal activity of essential oil from Ocimum gratissimum was investigated in order to evaluate its efficacy against Candida albicans, Candida krusei, Candida parapsilosis, and Candida tropicalis . Transmission and scanning electron microscopy and negative staining in light microscopy were performed to reveal the effects of the essential oil on the morphology of these yeasts . Determination of minimal inhibitory concentrations and time-kill curves demonstrated that the essential oil showed fungicidal activity against all of the Candida species studied . Analysis of the ultrastructure of the yeast cells revealed changes in the cell wall and in the morphology of some subcellular organelles . Bud formation in the yeasts was impaired in treated cells . The essential oil of O . gratissimum is a potential candidate as a phytotherapeutic agent in some fungal diseases and for the control of fungi in the environment. Appl Environ Microbiol, 2004 Aug, 70(8), 4872 - 9 Cloning and characterization of three fatty alcohol oxidase genes from Candida tropicalis strain ATCC 20336; Eirich LD et al.; Candida tropicalis (ATCC 20336) converts fatty acids to long-chain dicarboxylic acids via a pathway that includes among other reactions the oxidation of omega-hydroxy fatty acids to omega-aldehydes by a fatty alcohol oxidase (FAO) . Three FAO genes (one gene designated FAO1 and two putative allelic genes designated FAO2a and FAO2b), have been cloned and sequenced from this strain . A comparison of the DNA sequence homology and derived amino acid sequence homology between these three genes and previously published Candida FAO genes indicates that FAO1 and FAO2 are distinct genes . Both genes were individually cloned and expressed in Escherichia coli . The substrate specificity and K(m) values for the recombinant FAO1 and FAO2 were significantly different . Particularly striking is the fact that FAO1 oxidizes omega-hydroxy fatty acids but not 2-alkanols, whereas FAO2 oxidizes 2-alkanols but not omega-hydroxy fatty acids . Analysis of extracts of strain H5343 during growth on fatty acids indicated that only FAO1 was highly induced under these conditions . FAO2 contains one CTG codon, which codes for serine (amino acid 177) in C . tropicalis but codes for leucine in E . coli . An FAO2a construct, with a TCG codon (codes for serine in E . coli) substituted for the CTG codon, was prepared and expressed in E . coli . Neither the substrate specificity nor the K(m) values for the FAO2a variant with a serine at position 177 were radically different from those of the variant with a leucine at that position. Appl Microbiol Biotechnol . 2004 Jul 31; {Epub ahead of print} Biosurfactant from Lactococcus lactis 53 inhibits microbial adhesion on silicone rubber; Rodrigues L et al.; The ability of biosurfactant obtained from the probiotic bacterium Lactococcus lactis 53 to inhibit adhesion of four bacterial and two yeast strains isolated from explanted voice prostheses to silicone rubber with and without an adsorbed biosurfactant layer was investigated in a parallel-plate flow chamber . The microbial cell surfaces and the silicone rubber with and without an adsorbed biosurfactant layer were characterized using contact-angle measurements . Water contact angles indicated that the silicone-rubber surface with adsorbed biosurfactant was more hydrophilic (48 degrees ) than bare silicone rubber (109 degrees ) . The results showed that the biosurfactant was effective in decreasing the initial deposition rates of Staphylococcus epidermidis GB 9/6 from 2,100 to 220 microorganisms cm(-2) s(-1), Streptococcus salivarius GB 24/9 from 1,560 to 137 microorganisms cm(-2) s(-1), and Staphylococcus aureus GB 2/1 from 1,255 to 135 microorganisms cm(-2) s(-1), allowing for a 90% reduction of the deposition rates . The deposition rates of Rothia dentocariosa GBJ 52/2B, Candida albicans GBJ 13/4A, and Candida tropicalis GB 9/9 were far less reduced in the presence of the biosurfactant as compared with the other strains . This study constitutes a step ahead in developing strategies to prevent microbial colonization of silicone-rubber voice prostheses. Antimicrob Agents Chemother, 2004 Aug, 48(8), 3107 - 11 In vitro activities of ravuconazole and four other antifungal agents against fluconazole-resistant or -susceptible clinical yeast isolates; Cuenca-Estrella M et al.; The activities of ravuconazole and four other antifungal agents were tested against a collection of 1,796 clinical yeast isolates, including fluconazole-susceptible and -resistant strains . Ravuconazole was active against the majority of fluconazole-resistant isolates; but for 102 of 562 (18%) resistant isolates, mainly Candida tropicalis, Candida glabrata, and Cryptococcus neoformans, ravuconazole MICs were > or =1 microg/ml. Pediatr Infect Dis J, 2004 Jul, 23(7), 635 - 41 Risk factors for disseminated candidiasis in children with candidemia; Zaoutis TE et al.; OBJECTIVE: To determine the risk factors for disseminated infection in hospitalized children with candidemia . METHODS: We performed a nested case-control study within a cohort of hospitalized children with candidemia . The cohort was defined by all patients with positive blood cultures for Candida species in a large, urban, academic, tertiary care children's hospital from 1998 to 2001 . Cases were patients with clinical, microbiologic or radiographic evidence of disseminated candidiasis . Controls were patients with no evidence of disseminated candidiasis . RESULTS: Among 168 total children with candidemia, the median age was 3.5 years (interquartile range, 0.6-14.3) . There were 189 episodes of candidemia . Candida species included:Candida albicans (41%), Candida parapsilosis (24%), Candida glabrata (13%) and Candida tropicalis (9%) . The most common underlying diagnoses were oncologic (24%), gastrointestinal (15%) and cardiac (10%) diseases . Eighty-nine patients (53%) were admitted to an intensive care unit, 46 (27%) to a general pediatric or surgical ward and 33 (20%) to the oncology ward.Of the 168 patients with candidemia, 153 were included in the analysis of risk factors for disseminated candidiasis . Of 153 (17%) patients, 26 had evidence of organ dissemination . Organ involvement was most commonly identified in the lung (58%), followed by the liver (23%), kidney (16%), brain (12%), spleen (8%), eye (8%) and heart (8%) . Eight of the 26 patients had evidence of dissemination to more than 1 organ . Independent risk factors for disseminated candidiasis were persistently positive blood cultures for Candida (>3 days) with a central venous catheter in place (odds ratio, 3.0; 95% confidence interval, 1.2, 7.8; P = 0.02) and immunosuppression (odds ratio, 2.9; 95% confidence interval, 1.2, 7.0; P = 0.02) . CONCLUSIONS: Prolonged duration of candidemia with a central venous catheter in place and immunosuppression were independent risk factors for disseminated candidiasis in children with candidemia . Furthermore review of the epidemiology of candidemia at our institution revealed a heterogeneous population of children at risk for candidemia and a predominance of non-albicans species as the cause of these infections . Future studies are needed to determine the extent of evaluation needed for detecting dissemination among children with candidemia and to explore interventions for its prevention. Enferm Infecc Microbiol Clin, 2004 Jun-Jul, 22(6), 328 - 31 {In vitro activity of caspofungin against fluconazole-resistant Candida isolates from patients with HIV infection}; Ortiz de la Tabla-Ducasse V et al.; INTRODUCTION: Antifungal therapy for mucosal candidiasis caused by fluconazole-resistant Candida species is problematic . The aim of this study was to investigate the in vitro activity of caspofungin against Candida strains with reduced susceptibility to fluconazole isolated from HIV-infected patients . METHODS: The in vitro activity of caspofungin was assessed in 28 fluconazole-resistant Candida isolates obtained from the oral cavity of a cohort of 174 consecutive HIV-infected patients . Minimum inhibitory concentrations (MICs) were determined by a standardized broth microdilution method, as recommended by the NCCLS . RESULTS: Overall, caspofungin MICs ranged from < or = 0.06 microg/ml to 1 microg/ml . MICs at which 50% (MIC50) and 90% (MIC90) of isolates were inhibited were 0.25 microg/ml and 0.5 microg/ml, respectively . MICs ranged from < or = 0.06 microg/ml to 0.5 microg/ml for Candida albicans (n = 11), and < 0.06 microg/ml to 1 microg/ml or Candida glabrata (n = 11) . MICs for the two strains of Candida krusei were 0.125 microg/ml and 1 microg/ml . The range of MICs for Candida tropicalis and Candida inconspicua strains was 0.25 microg/ml to 0.5 microg/ml . CONCLUSION: Caspofungin was very active in vitro against a variety of fluconazole-resistant Candida strains recovered from a clinical cohort of HIV-infected patients . The MIC50 values and MIC ranges were slightly higher for Candida glabrata than for Candida albicans. J Microbiol Immunol Infect, 2004 Jun, 37(3), 157 - 63 Hemophagocytic syndrome: a review of 18 pediatric cases; Chen CJ et al.; This retrospective study included 18 pediatric cases (median age, 3 years) with pathologically proved hemophagocytic syndrome (HPS) from a single institution during 1992 and 2001 . There were 9 males and 9 females . Prolonged fever, cytopenia, liver dysfunction and hepatomegaly were the most common features at presentation . Sixteen (88.9%) cases were previously healthy . The case fatality rate was 61.1%, and all fatal cases died within 2 months of disease onset . The infectious agents associated with HPS were identified in 11 cases (61.1%), and 8 (72.7%) of them had evidence of Epstein-Barr virus (EBV) infection or reactivation . Underlying immunologic disorder or neoplastic disease was identified in 11.1% of the cases . Children less than 3 years of age with HPS were more vulnerable to neutropenia-associated bloodstream infection (85.7% vs 27.3%; p=0.025) . Pseudomonas aeruginosa (3) and Candida tropicalis (2) were the 2 most commonly isolated pathogens . Regarding specific management of HPS, intravenous immunoglobulin and steroids were the first-line agents and were administered in 16 cases and 11 cases, respectively, while etoposide was administered in 5 refractory cases during the late phase of disease . Most HPS occurred in previously healthy children, and a substantial proportion of cases rapidly progressed to death . Most cases were associated with viral infection, particularly EBV, and young children tended to develop neutropenia-associated bacteremia during the active phase of the disease. Pediatr Crit Care Med, 2004 Jul, 5(4), 369 - 74 Candidemia in a pediatric intensive care unit; Singhi SC et al.; OBJECTIVE: To examine the incidence, epidemiology, and clinical characteristics of candidemia in a pediatric intensive care unit . DESIGN: Retrospective cohort study . SETTING: Pediatric intensive care unit of a tertiary care teaching and referral hospital in north India . SUBJECTS: All patients with candidemia from March 1993 to December 1996 . INTERVENTIONS: Patient-related data were analyzed to study candidemia in relation to reason for fungal culture, underlying medical conditions, predisposing factors, Candida isolates, antimicrobial and antifungal treatment, and deaths . MEASUREMENTS AND MAIN RESULTS: Sixty-four patients with candidemia were identified . The Candida species isolated were Candida tropicalis (48.4%), C . albicans (29.7%), C . guillermondii (14.1%), C . krusei (6.3%), and C . glabrata (1.6%) . Thirty-three patients were detected by a high-risk surveillance blood culture, whereas 31 patients were detected while undergoing septic workup . Sixteen (25%) patients were asymptomatic; they recovered without any antifungal therapy and without any sequelae . Of 48 symptomatic patients, 11 died before institution of antifungal therapy; 37 received oral itraconazole (10 mg.kg(-1).day(-1)) . Seven (19%) of these 37 patients died . Those who recovered had sterile culture on average by day 14 (range, 4-30) and received the antifungal therapy on average for 24 days (range, 9-42 days) . Overall mortality rate was 28.1%, and bivariate analysis showed significant association with Pediatric Risk of Mortality score (p =.0001), presence of symptoms (p =.003), isolation of nonalbicans Candida in general (p =.04) and C . tropicalis specifically (p =.001), and failure to give presumptive antifungal therapy (p =.055) . On multivariate analysis, Pediatric Risk of Mortality score and isolation of C . tropicalis were the only significant predictors of mortality . CONCLUSIONS: Nonalbicans Candida accounted for 70% of candidemia in a pediatric intensive care unit . High-risk surveillance blood cultures aided diagnosis in about half the patients . Severity of illness and isolation of C . tropicalis were significant predictors of mortality. Asian Cardiovasc Thorac Ann, 2004 Jun, 12(2), 95 - 8 Fungal endocarditis: an autopsy study; Challa S et al.; Between 1990 and 2002, 237 hearts were examined at autopsy, including 16 with infective endocarditis; 6 showed fungal endocarditis . The preceding pathology was chronic rheumatic heart disease in 2 patients, one of whom had undergone double valve replacement, 2 patients had been treated for acute lymphoblastic leukemia, and one had protein-energy malnutrition . The underlying cause was unknown in one case . The organisms isolated were Aspergillus in 3 patients, Zygomycota in 1, Candida in 1, and both Candida tropicalis and Aspergillus in 1 patient . Immunosuppressed states are a cause of fungal endocarditis in India, although chronic rheumatic heart disease is the preceding pathology in the majority of patients. Biotechnol Lett, 2004 Apr, 26(8), 623 - 7 Increased xylitol production rate during long-term cell recycle fermentation of Candida tropicalis; Kim TB et al.; Long-term cell recycle fermentations of Candida tropicalis were performed over 14 rounds of fermentation . The average xylitol concentrations, fermentation times, volumetric productivities and product yields for 14 rounds were 105 g l(-1), 333 h, 4.4 g l(-1) h(-1) and 78%, respectively, in complex medium; and 110 g l(-1), 284 h, 5.4 g l(-1) h(-1) and 81%, respectively, in a chemically defined medium . These productivities were 1.7 and 2.4 times those with batch fermentation in the complex and chemically defined media, respectively . The xylitol yield from xylose with cell recycle fermentation using the chemically defined medium was 81% (w/w), which was 7% greater than the xylitol yield with batch fermentation (74%); both modes of fermentation gave the same yield using the complex medium . These results suggest that the chemically defined medium is more suitable for production of xylitol than complex medium. Saudi Med J, 2004 May, 25(5), 566 - 9 Distribution of Candida species among bloodstream isolates; Al-Jasser AM et al.; OBJECTIVE: To identify the distribution of Candida species causing bloodstream infections . METHODS: This study was conducted at the Armed Forces Hospital, Riyadh, Kingdom of Saudi Arabia . All cases of candidemia from the period 1996 through to 2002 were retrospectively identified through the records from the Department of Clinical Microbiology . RESULTS: Two hundred and ninety-four candidemic episodes were identified, 176 (59.9%) occurred in the intensive care units (ICUs), 32 (10.9%) medical, 30 (10.2%) surgical wards, 24 (8%) from patients with hematologic malignancies and 15 (5%) from pediatric wards . Candida albicans (C . albicans) was the most frequently isolated species with 149 (50.7%) cases, followed by Candida tropicalis (C . tropicalis) 61 (20.7%), Candida parapsilosis 32 (10.9%), Candida krusei (C . krusei) 23 (7.8%) and Candida glabrata 21 (7.1%) . Other species were not common . There is an increase in the proportion of non C . albicans species as the causative agents of candidemia . In certain clinical settings, non C . albicans species predominate as in the Adult General Intensive Care Unit with C . tropicalis as the most common . While in patients with hematologic malignancies, C . krusei species is the most common . CONCLUSION: These findings reinforce the need for continued and active surveillance programs to address the changes in the species distribution among candidal bloodstream isolates which will help to develop effective, preventive and therapeutic strategies. Mycopathologia, 2004 Feb, 157(2), 145 - 53 Anti-Candida and mode of action of two newly synthesized polymers: a modified poly (methylmethacrylate-co-vinylbenzoylchloride) and a modified linear poly (chloroethylvinylether-co-vinylbenzoylchloride) with special reference to Candida albicans and Candida tropicalis; Mahmoud YA et al.; Polymeric antimicrobial agents represent a new and important direction that is developing in the field of antimicrobial agents . Antimicrobial activity of two newly synthesized polymers: a modified poly (methylmethacrylate-co-vinylbenzoylchloride) and a modified linear poly (chloroethylvinylether-co-vinylbenzoylchloride) have been investigated and found to be active . Both polymers have showed a broad antimicrobial activity against C . albicans and C . tropicalis . Minimal inhibitory concentrations (MIC's) for poly (methylmethacrylate-co-vinylbenzoyl chloride) were 100, 75 and 100 microg/ml in case of C . albicans (ATCC 2091), C . albicans (SC5314) and C . tropicalis, respectively . However, polycholoroethylvinylether-covinylbenzoylchloride inhibited C . albicans (ATCC 2091), C . albicans (SC5314) and C . tropicalis with minimum inhibitory concentration values (MIC's) of 150 microg/ml against the three tested Candida strains . Mode of action studies of both polymers on the medically important yeasts, C . albicans and C . tropicalis revealed that poly (methylmethacrylate-co-vinylbenzoylchloride) induced cytotoxicity, DNA damage, and altered cell permeability and morphology, which was manifested as aggregated and swollen yeast cells (C . albicans ATCC 2091) by fluorescent microscopy examination . Poly (chloroethylvinylether-co-vinylbenzoylchloride) increased cell permeability, and respiration for C . albicans and C . tropicalis . The tested polymers at 50 microg/ml had pronounced effects on C . albicans and C . tropicalis cell wall phosphopeptidomannane, proteins, sugars and phosphorus . Generally, the two polymers proved effective against the tested microorganisms, but growth inhibitory effect varied according to the composition of the polymer active group . Many investigators consider polymeric antimicrobial agents as a potential new approach for enhancing the efficiency of some existing antimicrobial agents, including prolonged activity, reduce their toxicity, as well as reduce the environmental issues associated with product use. Int J Infect Dis, 2004 May, 8(3), 180 - 6 The predictors of outcome in immunocompetent patients with hematogenous candidiasis; Safdar A et al.; OBJECTIVE: Clinical parameters that predict outcome in non-immunosuppressed candidemic patients are not fully understood . METHODS: Eighty-one consecutive episodes of candidemia were retrospectively evaluated in 75 patients during 1998-2000 . RESULTS: Infection due to Candida albicans was common (n = 30; 37%) followed by Candida glabrata (n = 25; 31%), Candida parapsilosis (n = 14; 17%), Candida tropicalis (n = 6; 7%), Candida krusei (n = 5; 6%), and Candida lusitaniae (n = 1; 1%) . Among 70 evaluable patients, 31 (44%) had fungemia-associated mortality; advanced age (P < 0.004), underlying malignancy (P < 0.025), coronary artery disease (P < 0.01), and concurrent non-Candida species fungal infection (P < 0.047) were significant prognosticators of compromised short-term survival by multivariate analysis . Mortality was higher in patients with Candida glabrata (60%) and C . tropicalis (75%) infection compared to 44% deaths in individuals with C . albicans infection (P > 0.1) . 11/25 (44%) of non-immunocompromised individuals died and 20/45 (44%) immunosuppressed patients succumbed to fungemia: persistent vs . non-persistent (< 3 days) Candida bloodstream invasion, neutropenia, diabetes mellitus, renal insufficiency, prior antimicrobial therapy, cirrhosis of liver, abdomino-pelvis surgery, and critical-care-unit vs . non critical-care-unit admission did not significantly impact outcome in either group . All 11 infants, including nine with prematurity, survived Candida species bloodstream infection (P < 0.025) . CONCLUSIONS: Short-term mortality in candidemic non-immunocompromised patients was comparable to fungemia-associated deaths in immunosuppressed patients . Ischemic heart disease has appeared as a new predictor of unfavorable outcome in patients with hematogenous candidiasis. Environ Pollut, 1993, 80(1), 41 - 4 Susceptibility of different yeast species to environmental toxic metals; Berdicevsky I et al.; The purpose of the study reported here was to investigate the relative resistance of yeast species to various metallic and metalloid ions, with a view to gaining more knowledge on this subject, as resistant species may become dominant in habitats contaminated with the relevant metals . Saccharomyces cerevisiae, Candida albicans and Candida tropicalis were grown in media containing different concentrations of mercury (as HgCl(2)), cadmium (as CdCl(2)), lead (as Pb(CH(3)COO)(2)), arsenic (as Na(2)HAsO(4)) and selenium (as Na(2)SeO(3)) for various intervals . Invariably, the two Candida species turned out to be more resistant to all the metals studied than S . cerevisiae . The metal showing the highest toxicity for these species was mercury, with cadmium being the second, lead, the third and arsenic and selenium being the least toxic elements . Strains showing resistance to mercury were isolated, even in the case of S . cerevisiae. J Clin Microbiol, 2004 Apr, 42(4), 1716 - 8 Quality control limits for voriconazole disk susceptibility tests on Mueller-Hinton agar with glucose and methylene blue; Pfaller MA et al.; An international collaborative study was performed in order to propose quality control limits for voriconazole disk diffusion tests on Mueller-Hinton agar with 2% glucose and 0.5 micro g of methylene blue per ml . The supplement may be added to the agar before autoclaving, or Mueller-Hinton agar plates may be flooded with a glucose-methylene blue solution . Replicate tests on both types of agar plates with 1- micro g voriconazole disks generated data to propose zone size limits for tests of Candida parapsilosis ATCC 22019 (28 to 37 mm), Candida albicans ATCC 90028 (31 to 42 mm), and Candida krusei ATCC 6258 (16 to 25 mm) . Candida tropicalis ATCC 750 was not useful for this purpose. Biotechnol Lett, 2004 Feb, 26(4), 2065 - 9 Production of xylitol from Candida tropicalis by using an oxidation-reduction potential-stat controlled fermentation; Sheu DC et al.; An on-line device, ORP (oxidation-reduction potential)-stat, was used to control glucose-feeding for enhancing xylitol conversion from D-xylose during an oxygen-limited fermentation by Candida tropicalis . The fermentation was carried out in a 5 l jar fermenter . After glucose in the medium was depleted, a switching to a limited aeration and feeding glucose controlled by ORP-stat was performed . The maximum xylitol yield was obtained under a condition at an ORP of - 180 mV and at an aeration rate of 0.2 l min(-1). J Biol Chem, 2004 Jun 4, 279(23), 24666 - 72 Epub 2004 Mar 29. A two-domain structure of one subunit explains unique features of eukaryotic hydratase 2; Koski MK et al.; 2-Enoyl-CoA hydratase 2, a part from multifunctional enzyme type 2, hydrates trans-2-enoyl-CoA to 3-hydroxyacyl-CoA in the (3R)-hydroxy-dependent route of peroxisomal beta-oxidation of fatty acids . Unliganded and (3R)-hydroxydecanoyl coenzyme A-complexed crystal structures of 2-enoyl-CoA hydratase 2 from Candida tropicalis multifunctional enzyme type 2 were solved to 1.95- and 2.35-A resolution, respectively . 2-Enoyl-CoA hydratase 2 is a dimeric, alpha+beta protein with a novel quaternary structure . The overall structure of the two-domain subunit of eukaryotic 2-enoyl-CoA hydratase 2 resembles the homodimeric, hot dog fold structures of prokaryotic (R)-specific 2-enoyl-CoA hydratase and beta-hydroxydecanoyl thiol ester dehydrase . Importantly, though, the eukaryotic hydratase 2 has a complete hot dog fold only in its C-domain, whereas the N-domain lacks a long central alpha-helix, thus creating space for bulkier substrates in the binding pocket and explaining the observed difference in substrate preference between eukaryotic and prokaryotic enzymes . Although the N- and C-domains have an identity of <10% at the amino acid level, they share a 50% identity at the nucleotide level and fold similarly . We suggest that a subunit of 2-enoyl-CoA hydratase 2 has evolved via a gene duplication with the concomitant loss of one catalytic site . The hydrogen bonding network of the active site of 2-enoyl-CoA hydratase 2 resembles the active site geometry of mitochondrial (S)-specific 2-enoyl-CoA hydratase 1, although in a mirror image fashion . This arrangement allows the reaction to occur by similar mechanism, supported by mutagenesis and mechanistic studies, although via reciprocal stereochemistry. Antonie Van Leeuwenhoek, 2004 May, 85(4), 281 - 6 Characterization of a new xylitol-producer Candida tropicalis strain; Lopez F et al.; A xylitol-producer yeast isolated from corn silage and designated as ASM III was selected based on its outstanding biotechnological potential . When cultivated in batch culture mode and keeping the dissolved oxygen at 40% saturation, xylitol production was as high as 130 g l(-1) with a yield of 0.93 g xylitol g(-1) xylose consumed . A preliminary identification of the yeast was performed according to conventional fermentation and assimilation physiological tests . These studies were complemented by using molecular approaches based on PCR amplification, restriction-fragment length polymorphism analysis and sequencing of the rDNA segments: intergenic transcribed spacer (ITS) 1-5.8S rDNA-ITS 2, and D1/D2 domain of the 26S rRNA gene . Results from both the conventional protocols and the molecular characterization, and proper comparisons with the reference strains Candida tropicalis ATCC 20311 and NRRL Y-1367, led to the identification of the isolate as a new strain of C . tropicalis. Haematologica, 2004 Mar, 89(3), 378 - 80 Clinical significance of breakthrough fungemia caused by azole-resistant Candida tropicalis in patients with hematologic malignancies; Myoken Y et al.; A 5-year retrospective analysis of fungemia in patients with hematologic malignancies revealed that four patients, who received fluconazole and itraconazole during neutropenia, developed breakthrough candidemia due to azole-resistant Candida tropicalis isolates . This observation suggests that causative organisms of candidemia in neutropenic patients receiving azoles should be suspected of being azole-resistant. J Clin Microbiol, 2004 Mar, 42(3), 1024 - 9 Human beta-defensins 2 and 3 demonstrate strain-selective activity against oral microorganisms; Joly S et al.; Human beta-defensins 2 and 3 (HBD-2 and HBD-3) are inducible peptides present at sites of infection in the oral cavity . A few studies have reported broad-spectrum antimicrobial activity for both peptides . However, no comprehensive study has thoroughly investigated their potential against oral pathogens . The purpose of this study was to test the effectiveness of HBD-2 and HBD-3 against a collection of oral organisms (Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis, Peptostreptococcus micros, Actinomyces naeslundii, Actinomyces israelii, Streptococcus sanguis, Streptococcus mutans, Candida tropicalis, Candida parapsilosis, Candida krusei, Candida glabrata, and Candida albicans) . Radial diffusion assays were used to test HBD-2 and HBD-3 activities against at least three strains of each species . There was significant variability in MICs, which was strain specific rather than species specific . MICs ranged from 3.9 to >250 micro g/ml for HBD-2 and from 1.4 to >250 micro g/ml for HBD-3 . HBD-3 demonstrated greater antimicrobial activity and was effective against a broader array of organisms . Overall, aerobes were 100% susceptible to HBD-2 and HBD-3, whereas only 21.4 and 50% of the anaerobes were susceptible to HBD-2 and HBD-3, respectively . HBD-2 and HBD-3 also demonstrated strain-specific activity against the Candida species evaluated . Interestingly, an association between HBD-2 and HBD-3 activities was noted . This suggests that the two peptides may have similar mechanisms yet utilize distinct pathways . The lack of activity against specific anaerobic strains and Candida warrants further investigation of the potential resistance mechanisms of these organisms . Finally, the significant variability between strains underlies the importance of testing multiple strains when evaluating activities of antimicrobial peptides. Antimicrob Agents Chemother, 2004 Mar, 48(3), 873 - 8 In vitro activities of 3-(halogenated phenyl)-5-acyloxymethyl- 2,5-dihydrofuran-2-ones against common and emerging yeasts and molds; Buchta V et al.; Three 3-(halogenated phenyl)-5-acyloxymethyl-2,5-dihydrofuran-2-ones were evaluated for activity against 191 strains of common and emerging yeasts and Aspergillus species by the broth microdilution test performed according to NCCLS guidelines . The furanone derivatives displayed broad-spectrum in vitro activity against potentially pathogenic yeasts and molds, especially Aspergillus spp . (MIC <or= 2.0 microg/ml) and fluconazole-resistant yeast isolates, including Candida glabrata and Saccharomyces cerevisiae . The 4-bromophenyl derivative was the most effective derivative against the majority of species tested, except for the Candida tropicalis and C . glabrata strains, which were more susceptible to the 3-chlorophenyl derivative . The 3,4-dichlorophenyl derivative possessed a lesser in vitro antifungal effect . The potential of further experiments on animal infection and clinical studies is supported by the relatively low cytotoxicity and acute toxicity of the 4-bromophenyl compound . Thus, the halogenated 3-phenyl-5-acyloxymethyl derivatives of 2,5-dihydrofuran-2-one represent a novel, promising group of compounds with significant activity against relevant opportunistic fungi that are pathogenic to humans. Antimicrob Agents Chemother, 2004 Mar, 48(3), 774 - 9 Quantitative evaluation of the therapeutic effects of antibiotics using silkworms infected with human pathogenic microorganisms; Hamamoto H et al.; The injection of bacteria (Staphylococcus aureus, Stenotrophomonas maltophilia) or true fungi (Candida albicans, Candida tropicalis) that are pathogenic to humans into the silkworm hemolymph leads to death of the larvae within 2 days . Antibiotics used for clinical purposes have therapeutic effects on silkworms infected with these pathogens . The 50% effective doses obtained by injection into the silkworm hemolymph are consistent with those reported for mice . Injection of vancomycin and kanamycin into the silkworm hemolymph was effective, but oral administration was not . Chloramphenicol, which is effective by oral administration, appeared in the silkworm hemolymph soon after injection into the midgut, whereas vancomycin did not . Isolated midgut membranes were impermeable to vancomycin . Thus, the ineffectiveness of oral administration of vancomycin to silkworms is due to a lack of intestinal absorption. Folia Microbiol (Praha), 2003, 48(5), 679 - 81 Antifungal activity of some bis-5-methylbenzimidazole compounds; Kucukbay H et al.; Twenty bis-5-methylbenzimidazole compounds were evaluated for their in vitro antifungal activity against Candida albicans and Candida tropicalis . Except for three all compounds exhibited an antifungal activity against these yeasts over a range of the minimum inhibitory concentration (MIC) between 25 and 800 mg/L. Biotechnol Lett, 2003 Dec, 25(24), 2085 - 8 Xylitol production by Candida tropicalis in a chemically defined medium; Kim TB et al.; A chemically defined medium that included urea (5 g l(-1)) as a nitrogen source and various vitamins was substituted for a complex medium containing yeast extract (10 g l(-1)) in the production of xylitol by Candida tropicalis . In a fed-batch culture with the chemically defined medium, 237 g xylitol l(-1) was produced from 270 g xylose l(-1) after 120 h . The volumetric rate of xylitol production and the xylitol yield from xylose were 2 g l(-1) h(-1) and 89%, respectively . These values were about 5% lower and 4% higher, respectively, than those obtained using the complex medium . These results indicate that xylitol can be produced effectively in a chemically defined medium. Biotechnol Lett, 2003 Dec, 25(24), 2065 - 9 Production of xylitol from Candida tropicalis by using an oxidation-reduction potential-stat controlled fermentation; Sheu DC et al.; An on-line device, ORP (oxidation-reduction potential)-stat, was used to control glucose-feeding for enhancing xylitol conversion from D-xylose during an oxygen-limited fermentation by Candida tropicalis . The fermentation was carried out in a 5 l jar fermenter . After glucose in the medium was depleted, a switching to a limited aeration and feeding glucose controlled by ORP-stat was performed . The maximum xylitol yield was obtained under a condition at an ORP of -180 mV and at an aeration rate of 0.2 l min(-1). Diagn Microbiol Infect Dis, 2004 Jan, 48(1), 33 - 7 Distribution and antifungal susceptibility of Candida species causing candidemia from 1996 to 1999; Cheng MF et al.; Susceptibilities to amphotericin B and fluconazole of 383 Candida species isolated from blood were determined . Candida albicans was the most common species (55.6%), followed by Candida parapsilosis (17.5%), Candida tropicalis (16.5%), Candida glabrata (5.2%), Candida guilliermondii (2.3%), and others (2.9%) . All but three isolates, Candida ciferrii, C . tropicalis, and C . glabrata, one each, were susceptible to amphotericin B . A total of 367 (95.8%) and 15 (4.2%) isolates were susceptible and susceptible-dose dependent to fluconazole, respectively . Only one isolate, a C . glabrata, was resistant to fluconazole . Few patients (13%) having prior fluconazole treatments may explain the low rate of resistance to fluconazole in this study. Infect Control Hosp Epidemiol, 2004 Jan, 25(1), 60 - 4 Susceptibilities of Candida species to amphotericin B and fluconazole: the emergence of fluconazole resistance in Candida tropicalis; Yang YL et al.; OBJECTIVE: To determine the susceptibilities of Candida species isolated from Taiwan to amphotericin B and fluconazole . DESIGN: Prospective surveillance study . METHODS: Each hospital was asked to submit up to 10 C . albicans and 40 non-albicans Candida species during the collection period, from April 15 to June 15, 1999 . One isolate was accepted from each episode of infection . The broth microdilution method was used to determine susceptibilities to amphotericin B and fluconazole . RESULTS: Only 3 of 632 isolates, one each of C . famata, C . krusei, and C . tropicalis, were resistant to amphotericin B . A total of 53 (8.4%) of 632 clinical yeast isolates, consisting of 4% C . albicans, 8% C . glabrata, 15% C . tropicalis, and 70% C . krusei, were resistant to fluconazole . In contrast, no C . parapsilosis isolate was resistant to fluconazole . Isolates from tertiary-care medical centers had higher rates of resistance to fluconazole than did those from regional and local hospitals (11.4% vs 6.6%) . Isolates from different sources showed different levels of susceptibility to fluconazole . All of the isolates with the exception of C . tropicalis and C . krusei isolated from blood were susceptible to fluconazole . A pattern of co-resistance to both amphotericin B and fluconazole was observed . CONCLUSIONS: Non-albicans Candida species had higher rates of resistance to fluconazole than did C . albicans (44 of 395 {11.2%} vs 9 of 237 {3.8%}; P = .002) . The increasing rate of fluconazole resistance in C . tropicalis (15%) is important because C . tropicalis is one of the most commonly isolated non-albicans Candida species. Chembiochem, 2004 Feb 6, 5(2), 206 - 13 Chimeras of the homing endonuclease PI-SceI and the homologous Candida tropicalis intein: a study to explore the possibility of exchanging DNA-binding modules to obtain highly specific endonucleases with altered specificity; Steuer S et al.; Homing endonucleases are extremely specific endodeoxyribonucleases . In vivo, these enzymes confer mobility on their genes by inducing a very specific double-strand cut in cognate alleles that lack the cooling sequence for the homing endonuclease; the cellular repair of the double-strand break with the endonuclease-containing allele as a template leads to integration of the endonuclease gene, completing the homing process . As a result of their extreme sequence specificity, homing endonucleases are promising tools for genome engineering . For this purpose, it is desirable to design enzymes with defined new specificities . To analyse which DNA-binding elements are potential candidates for use in the design of enzymes with modified or even new specificity, we produced several chimeric proteins derived from the Saccharomyces cerevisiae VMA1 intein (PI-SceI) and the related Candida tropicalis VMA1 intein . Although the mature Candida intein is devoid of endonucleolytic activity, the exchange of two DNA-binding modules of PI-SceI with the homologous elements from the Candida intein results in an active endonuclease . The low sequence homology in these modules indicates that different protein-DNA contacts are responsible for the recognition of related DNA sequences . This flexibility in DNA recognition should, in principle, allow endonucleases to be produced with new specificities useful for genome engineering. J Clin Pathol, 2004 Feb, 57(2), 196 - 8 Polymicrobial candidaemia revealed by peripheral blood smear and chromogenic medium; Yera H et al.; Candida spp are the fourth most common group of nosocomial pathogens isolated from patients on medical, surgical, and intensive care wards . Polymicrobial candidaemia has rarely been described . The diagnosis of candidaemia from peripheral blood smears has not been widely reported . This report describes the case of a young woman suffering from Ewing's sarcoma who developed a syndrome of septic shock . Deep fungal infection was diagnosed from a systematic peripheral blood smear and yeasts were isolated within 24 hours . A subculture on CHROMagar Candida allowed the differentiation and presumptive identification of Candida tropicalis and Candida krusei . Species identification was confirmed by the ID 32C system . This report underlines the usefulness of peripheral blood smears in the diagnosis of fulminant deep fungal infections, and of a differential isolation medium in the rapid presumptive identification of clinically important yeast species from clinical samples . This medium is particularly useful for the detection of mixed fungal infections, allowing early and better adapted antifungal treatment. Appl Microbiol Biotechnol, 2004 May, 64(4), 531 - 6 Epub 2004 Jan 22. Heterologous expression of metabotropic glutamate receptor subtype 1 in Saccharomyces cerevisiae; Sugiyama K et al.; The upstream region of the isocitrate lyase gene (UPR-ICL) from the n-alkane-utilizing yeast Candida tropicalis serves as a useful promoter of gene expression in the yeast Saccharomyces cerevisiae . The production of rat metabotropic glutamate receptor 1alpha (mGluR1alpha), which belongs to the G-protein-coupled receptor (GPCR) family, was tested under the control of UPR-ICL . Expression of mGluR1alpha was found in recombinant clones and enhanced by replacing the signal sequence of mGluR1alpha with the corresponding region of the alpha-factor receptor (Ste2), which is a GPCR found in S . cerevisiae . Moreover, the membrane fraction from a recombinant clone associated with Vesl-1S/Homer-1a protein binds the mGluR1alpha in rat cerebellum . These results suggest that the UPR-ICL-controlled gene expression system is useful for heterologous GPCRs in S . cerevisiae . Clin Infect Dis, 2004 Feb 1, 38(3), 311 - 20 Epub 2004 Jan 14. Epidemiology of candidemia in Swiss tertiary care hospitals: secular trends, 1991-2000; Marchetti O et al.; Candida species are among the most common bloodstream pathogens in the United States, where the emergence of azole-resistant Candida glabrata and Candida krusei are major concerns . Recent comprehensive longitudinal data from Europe are lacking . We conducted a nationwide survey of candidemia during 1991-2000 in 17 university and university-affiliated hospitals representing 79% of all tertiary care hospital beds in Switzerland . The number of transplantations and bloodstream infections increased significantly (P<.001) . A total of 1137 episodes of candidemia were observed: Candida species ranked seventh among etiologic agents (2.9% of all bloodstream isolates) . The incidence of candidemia was stable over a 10-year period . C . albicans remained the predominant Candida species recovered (66%), followed by C . glabrata (15%) . Candida tropicalis emerged (9%), the incidence of Candida parapsilosis decreased (1%), and recovery of C . krusei remained rare (2%) . Fluconazole consumption increased significantly (P<.001) . Despite increasing high-risk activities, the incidence of candidemia remained unchanged, and no shift to resistant species occurred. Indian J Pediatr, 2003 Nov, 70(11), 925 - 7 Candida tropicalis meningitis in a young infant; Ahuja SR et al.; Candida tropicalis is a rare species of Candida causing meningitis . The authors report a young infant who developed Candida tropicalis meningitis following a prolonged stay in a neonatal intensive care unit for respiratory distress and intra-cranial hemorrhage . The child was successfully treated with recommended doses of Amphotericin B and 5-flucytosine for eight weeks. J Antimicrob Chemother, 2004 Feb, 53(2), 283 - 9 Epub 2003 Dec 19. National surveillance of species distribution in blood isolates of Candida species in Japan and their susceptibility to six antifungal agents including voriconazole and micafungin; Takakura S et al.; OBJECTIVES: The aim of this study was to evaluate species distribution and antifungal susceptibility of Candida blood isolates in Japan . METHODS: In a 1 year surveillance programme, 535 Candida blood isolates were collected . Identification of species was followed by examination with the broth microdilution method, as described in NCCLS M27-A2, of antifungal susceptibility to six agents, including voriconazole and micafungin, with readings after 24 and 48 h of incubation . RESULTS: The overall species distribution was: 41% Candida albicans, 23% Candida parapsilosis, 18% Candida glabrata, 12% Candida tropicalis and 2% Candida krusei . The concentrations of fluconazole necessary to inhibit 90% of the isolates (MIC(90)) at 24/48 h were 0.25/1 mg/L for C . albicans, 0.5/2 mg/L for C . parapsilosis, 4/32 mg/L for C . glabrata and 4/>128 mg/L for C . tropicalis . Percentages of fluconazole resistance were 1.8% for C . albicans, 0.8% for C . parapsilosis, 5.2% for C . glabrata and 3.2% for C . tropicalis, taking the tendency of trailing growth of C . tropicalis into account . MIC(90) of voriconazole was 0.5 mg/L, although 35% of isolates less susceptible (>/=16 mg/L) to fluconazole showed resistance (>/=2 mg/L) . Micafungin was very active against all species (MIC(90), 0.03 mg/L) except for C . parapsilosis (MIC(90), 2 mg/L) . CONCLUSIONS: These data suggest that, in Japan, the species distribution of Candida bloodstream infections and the fluconazole resistance rate are similar to those reported previously in North America and Europe . Voriconazole and micafungin appear to have strong in vitro activity against Candida blood isolates, although continuing surveillance and further clinical research are needed. Gen Physiol Biophys, 2003 Jun, 22(2), 167 - 79 Hydroxylation of phenol to catechol by Candida tropicalis: involvement of cytochrome P450; Stiborova M et al.; Microsomal preparations isolated from yeast Candida tropicalis (C . tropicalis) grown on three different media with or without phenol were isolated and characterized for the content of cytochrome P450 (CYP) (EC 1.14.15.1) . While no CYP was detected in microsomes of C . tropicalis grown on glucose as the carbon source, evidence was obtained for the presence of the enzyme in the microsomes of C . tropicalis grown on media containing phenol . Furthermore, the activity of NADPH: CYP reductase, another enzyme of the microsomal CYP-dependent system, was markedly higher in cells grown on phenol . Microsomes of these cells oxidized phenol . The major metabolite formed from phenol by microsomes of C . tropicalis was characterized by UV/vis absorbance and mass spectroscopy as well as by the chromatographic properties on HPLC . The characteristics are identical to those of catechol . The formation of catechol was inhibited by CO, the inhibitor of CYP, and correlated with the content of cytochrome P450 in microsomes . These results, the first report showing the ring hydroxylation of phenol to catechol with the microsomal enzyme system of C . tropicalis, strongly suggest that CYP-catalyzed reactions are responsible for this hydroxylation . The data demonstrate the progress in resolving the enzymes responsible for the first step of phenol degradation by the C . tropicalis strain. Braz J Infect Dis, 2003 Dec, 7(6), 426 - 8 Septic arthritis as the first sign of Candida tropicalis fungaemia in an acute lymphoid leukemia patient; Vicari P et al.; Fungal infections caused by Candida species have increased in incidence during the past two decades in England, North America and Europe . Candidal arthritis is rare in patients who are not intravenous drug users or are who not using a prostheses . We report the case of a 24-year-old man with acute lymphoid leukemia, who developed Candida tropicalis arthritis during an aplastic period after chemotherapy . This is the eighth case described in the literature of C . tropicalis causing arthritis without intra-articular inoculation . We call attention to an unusual first sign of fungal infection: septic arthritis without intra-articular inoculation . However, this case differs from the other seven, since despite therapy a fast and lethal evolution was observed . We reviewed reported cases, incidence, risk factors, mortality and treatment of neutropenic patients with fungal infections. J Basic Microbiol, 2003, 43(6), 530 - 3 Identification of yeasts isolated from Nigerian sugar cane peels; Olasupo NA et al.; A pilot study was carried out to determine the yeast species associated with sugar cane peels recovered from three separate dumping sites found in Ojo, a typical market town in Nigeria . Species identities were determined by 26S ribosomal DNA (rDNA) sequencing . By using this method, a total of 6 different yeasts were identified, with Candida tropicalis, a recognised human yeast pathogen, being the species most frequently isolated from these dumping sites . The implications of this and other findings of the study, the apparent first of its kind, are discussed in detail. Int J Food Microbiol, 2003 Dec 31, 89(2-3), 275 - 80 Microbial modification of the texture of grated cassava during fermentation into akyeke; Obilie EM et al.; The traditional akyeke inoculum and fermenting akyeke, an indigenous cassava product, were investigated to identify microbial species responsible for the modification of cassava texture during fermentation . Both field and laboratory samples were examined and only some cultures isolated on Plate Count Agar and Malt Extract Agar were found to be capable of causing a softening of cassava tissue when plated directly on sterile cassava slices . The cassava tissue softening isolates on PCA were tentatively identified as Bacillus subtilis and isolates on MEA as Candida tropicalis and Zygosacchromyces florentinus . The population of B . subtilis in the laboratory sample of inoculum was found to be 2.4 x 10(9) cfu g(-1) and increased during dough fermentation from 1.1 x 10(7) to 3.5 x 10(9) cfu g(-1) after 96 h . C . tropicalis was present in the inoclum at 3.0 x 10(9) cfu g(-1) and increased during dough fermentation from 3.2 x 10(6) to 6.9 x 10(7) cfu g(-1) whilst Z . florentinus was present in the inoclum at 9.1 x 10(8) cfu g(-1) and increased from 8.1 x 10(5) to 7.5 x 10(6) cfu g(-1) during dough fermentation. Oral Microbiol Immunol, 2003 Dec, 18(6), 379 - 88 Rapid and unequivocal differentiation of Candida dubliniensis from other Candida species using species-specific DNA probes: comparison with phenotypic identification methods; Ellepola AN et al.; Candida dubliniensis is a recently described opportunistic pathogen which shares many phenotypic characteristics with Candida albicans but which has been reported to rapidly acquire resistance to azole antifungal drugs . Therefore, differentiation of C . dubliniensis from C . albicans becomes important to better understand the clinical significance and epidemiologic role of C . dubliniensis in candidiasis . We compared phenotypic methods for the differentiation of C . dubliniensis from C . albicans (i.e . the ability to grow at elevated temperatures, colony color on CHROMagar Candida medium, and carbohydrate assimilation patterns) to amplify the results of a polymerase chain reaction (PCR) assay using universal fungal primers to the internal transcribed spacer 2 (ITS2) region of rDNA and species-specific DNA probes in an enzyme immunoassay format (PCR-EIA) . DNA sequencing of the ITS1 rDNA region was also conducted . The C . dubliniensis ITS2 probe correctly identified all C . dubliniensis isolates without cross-reaction with any other Candida species tested (mean A(650 nm) +/- SE, C . dubliniensis probe with C . dubliniensis DNA, 0.372 +/- 0.01, n = 22; C . dubliniensis probe with other Candida species DNA, 0.001 +/- 0.02 n = 16, P < 0.001) . All other Candida species tested (C . albicans, Candida glabrata, Candida krusei, Candida parapsilosis, and Candida tropicalis) were also correctly identified by the PCR-EIA without any detectable cross-reactions among species . Phenotypically, C . dubliniensis isolates demonstrated an increased sensitivity to heat compared to C . albicans isolates . At 42 degrees C, only 50% of C . dubliniensis isolates grew compared to 73% of C . albicans isolates and, at 45 degrees C, 91% of C . dubliniensis isolates failed to grow compared to 64% of C . albicans isolates . C . albicans was more likely to demonstrate a dark green or blue green colony color on CHROMagar Candida medium obtained from Becton Dickinson (i.e . 100% of C . albicans isolates were dark green or blue green versus 64% of C . dubliniensis isolates) whereas no difference in the percentage of C . albicans or C . dubliniensis isolates producing dark green or blue green colony color was detected using CHROMagar Candida medium from Hardy Diagnostics (82% for both species) . The API 20C AUX carbohydrate assimilation system incorrectly identified C . dubliniensis as C . albicans in all but three cases: remaining isolates were misidentified as C . albicans/C . tropicalis, C . tropicalis/C . albicans, and Candida lusitaniae/C . albicans . In all, 82% of C . albicans isolates and 100% of C . dubliniensis isolates assimilated trehalose; the latter finding was opposite to that reported for C . dubliniensis in the API 20C AUX profile index . Xylose and alpha-methyl-D-glucoside assimilation, respectively, were negative for 100 and 95% of C . dubliniensis isolates and positive for 100 and 91% of C . albicans isolates, confirming earlier reports that assimilation results for xylose and alpha-methyl-D-glucoside may be helpful in the discrimination of these two species . However, conventional phenotypic species identification tests required days for completion, whereas the PCR-EIA could be completed