Candida tropicalis

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 in a matter of hours . In addition, identification of Candida species by ITS1 rDNA sequencing gave 100% correspondence to the results obtained by the PCR-EIA, confirming the specificity of the PCR-EIA method . These data indicate that although a combination of phenotypic methods may help differentiate C . dubliniensis from C . albicans to some extent, the PCR-EIA can provide a simple, rapid, and unequivocal identification of the most medically important Candida species in a single test.

J Med Microbiol, 2003 Dec, 52(Pt 12), 1071 - 6
Species identification of medically important fungi by use of real-time LightCycler PCR; Hsu MC et al.; Invasive fungal infection has become a major cause of morbidity and mortality in immunocompromised patients . Rapid identification of pathogenic fungi to species level is critical for disease treatment . A real-time LightCycler assay aiming at rapid detection and species identification of pathogenic fungi from clinical isolates was developed . Template DNAs of different species were amplified and detected in real time by employing SYBR Green fluorescent dye . The target sequences for species-level detection were located between the 18S and 28S rDNA . Seven fungal species encountered frequently in the clinical setting, Candida albicans, Candida glabrata, Candida krusei, Candida parapsilosis, Candida tropicalis, Candida guilliermondii and Cryptococcus neoformans, could be discriminated by species-specific primers and confirmed by melting-curve analyses . The range of linearity was from 1 ng to 1 pg (microl(-1) water) and the sensitivity was 1 pg fungal DNA microl(-1) . Identification by this real-time PCR method matched biochemical identification for all 58 clinical strains . Therefore, the method is simple, rapid and sensitive enough for detection and identification of several fungal species.

Microbiology, 2003 Nov, 149(Pt 11), 3239 - 46
Inhibition of adhesion of yeasts and bacteria by poly(ethylene oxide)-brushes on glass in a parallel plate flow chamber; Roosjen A et al.; Poly(ethylene oxide) (PEO)-brushes are generally recognized as protein-repellent surfaces, and although a role in discouraging microbial adhesion has been established for some strains and species, no study exists on the effects of PEO-brushes on a large variety of bacterial and yeast strains . In this paper, a PEO-brush has been covalently attached to glass and silica by reaction in a polymer melt . Subsequently, the presence of a PEO-brush was demonstrated using contact angle measurements, X-ray photoelectron spectroscopy and ellipsometry . For five bacterial (Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus salivarius, Escherichia coli and Pseudomonas aeruginosa) and two yeast strains (Candida albicans and Candida tropicalis), adhesion to PEO-brushes was compared with adhesion to bare glass in a parallel plate flow chamber . The initial deposition rates of Sta . epidermidis, Sta . aureus and Str . salivarius to glass were relatively high, between 2400 and 2600 cm(-2) s(-1), while E . coli and P . aeruginosa deposited much more slowly . The initial deposition rates of the yeasts to glass were 144 and 444 cm(-2) s(-1) for C . albicans GB 1/2 and C . tropicalis GB 9/9, respectively . Coating of the glass surface with a PEO-brush yielded more than 98 % reduction in bacterial adhesion, although for the more hydrophobic P . aeruginosa a smaller reduction was observed . For both yeast species adhesion suppression was less effective than for the bacteria and here too the more hydrophobic C . tropicalis showed less reduction than the more hydrophilic C . albicans . The PEO-brush had a thickness of 22 nm in water, as inferred from ellipsometry . Assuming that on bare glass the adhered micro-organisms are positioned only a few nanometers away from the surface and that the brush keeps them at a distance of 22 nm, it is calculated that the brush yields a sevenfold attenuation of the Lifshitz-Van der Waals attraction to the surface between the micro-organisms and the surface . Decreased Lifshitz-van der Waals attraction may be responsible for the suppression of the microbial adhesion observed.

J Microbiol Immunol Infect, 2003 Sep, 36(3), 187 - 91
Fluconazole resistance rate of Candida species from different regions and hospital types in Taiwan; Yang YL et al.; From April 15 to June 15, 1999, 581 clinical Candida isolates from 19 hospitals in Taiwan were collected and susceptibilities to fluconazole of these isolates were determined by a broth microdilution method . A total of 42 (7.2%) isolates were resistant to fluconazole . Isolates from medical centers had a higher resistance rate to fluconazole than those from regional hospitals (10.7% vs 4.9%) . Candida species isolated from different regions had different degrees of susceptibility to fluconazole . Approximately 2.5%, 6.5%, and 11.8% of Candida isolates from middle, north, and south regions, respectively, were resistant to fluconazole . The prevalence of the combination of Candida glabrata, Candida krusei, and Candida tropicalis infections were 44.5%, 49.8%, and 62.7% in middle, north, and south regions, respectively . There is an association between the rate of fluconazole resistance and the number of non-albicans Candida species collected from different regions and hospital types.

Saudi Med J, 2003 Oct, 24(10), 1060 - 3
Candidemia and the susceptibility pattern of Candida isolates in blood; Osoba AO et al.; OBJECTIVE: Candida species has become one of the most common blood isolates as well as one of the leading causes of nosocomial bloodstream infections . The purpose of our study was to determine the prevalence of Candida species among our bloodstream infecting organisms and the susceptibility pattern of the Candida isolates to antifungal agents . METHODS: A prospective study was carried out in the Division of Microbiology, King Khalid National Guard Hospital, Jeddah, Kingdom of Saudi Arabia of all positive blood cultures for Candida species . The study took place from 1st January 1998 to March 2002 . Identification and susceptibility pattern of isolates were determined by the Candifast technique to amphotericin B, fluconazole, nystatin, Flucytosine, econazole, ketoconazole and miconazole . RESULTS: Over a 2-year period, 17,916 blood cultures were performed in our hospital . There were 2,972 positive cultures, of which 83 (2.8%) patients had Candida species isolated from their bloodstream . Of these, 38 (46%) were Candida albicans (C.albicans) . The remaining 45 strains were made up of Candida tropicalis 9 (10.8%); Candida parapsilosis 9 (10.8%); Candida species 9 (10.8%); Candida guilliermondi 6 (7.2%); Candida krusei 5 (6%); Candida glabrata 4 (4.8%); Candida pseudotropicalis 2 (2.4%) and Trichosporon species 1 (1.2%) . All Candida species were susceptible to amphotericin B . However, only 18 (47%) out of 38 C.albicans were susceptible to fluconazole, while only 8 (17.7%) of 45 non-C.albicans strains were susceptible to this drug . CONCLUSION: The susceptibility of C.albicans to fluconazole in our hospital using the Candifast method is very low (47%) . These results need to be confirmed by carrying out the Etest or the NCCLS M27-A method to confirm the true susceptibilities of Candida strains in our locality.

Eur J Clin Microbiol Infect Dis, 2003 Nov, 22(11), 651 - 5 Epub 2003 Oct 17.
Voriconazole salvage treatment of invasive candidiasis; Ostrosky-Zeichner L et al.; Data on the salvage treatment of invasive candidiasis with voriconazole in 52 patients intolerant of other antifungal agents or with infection refractory to other antifungal agents were analyzed . Patients had received a mean of two previous antifungal agents (range, 1-4 agents), and 83% had received an azole . Manifestations of invasive candidiasis included candidemia (37%), disseminated disease (25%), and infection of other sites (38%) . The median duration of voriconazole therapy was 60 days (range, 1-314 days) . The overall rate of response was 56% (95%CI, 41-70), with the following response rates observed for individual Candida species: Candida albicans, 44% (20-70); Candida glabrata, 38% (14-68); Candida krusei, 70% (35-93); Candida tropicalis, 67% (30-93); and other Candida spp., 100% (40-100) . The response rate in patients who had failed previous azole therapy was 58% (42-73) . Common adverse events (~20%) included nausea and emesis, abnormal liver enzymes, and visual disturbances . Serious adverse events occurred in four patients, and nine patients died . Voriconazole has promise as a salvage agent for the treatment of invasive candidiasis, even in the settings of previous azole therapy and infection due to Candida krusei.

Eur J Clin Microbiol Infect Dis, 2003 Nov, 22(11), 693 - 6 Epub 2003 Oct 14.
Rapid identification of yeasts commonly found in positive blood cultures by amplification of the internal transcribed spacer regions 1 and 2; Li YL et al.; A multiplex PCR method using one universal and eight species-specific primers was developed to rapidly identify eight yeast species found in positive blood cultures . The species-specific primers were designed from the internal transcribed spacer regions 1 and 2 of the rRNA gene, whereas the universal primer was located at the 26S rRNA gene . The eight species were Candida albicans, Candida glabrata, Candida guilliermondii, Candida krusei, Candida lusitaniae, Candida parapsilosis, Candida tropicalis, and Cryptococcus neoformans . The PCR products (116 to 630 bp) were different in length and could be effectively separated and recognized by polyacrylamide gel electrophoresis . By testing 234 positive blood cultures (237 isolates), 234 (98.7%) isolates of the above eight species were correctly identified by the multiplex PCR . The present method is simple to perform and can be completed within 6 h.

J Clin Microbiol, 2003 Oct, 41(10), 4551 - 8
Contribution of the Platelia Candida-specific antibody and antigen tests to early diagnosis of systemic Candida tropicalis infection in neutropenic adults; Sendid B et al.; The Platelia Candida-specific antigen and antibody assays (Bio-Rad Laboratories) were used to test serial serum samples from seven neutropenic adult patients with hematological malignancies who had developed systemic Candida tropicalis infections . The diagnosis of candidiasis was based on a positive blood culture (all seven patients) and the isolation of C . tropicalis from a normally sterile site (six patients) . All patients received early antifungal therapy with amphotericin B and/or an azole derivative and had successful outcomes . When the combined assays were applied to sera collected at different time points before and after the first positive blood culture, all patients tested positive . In six patients, at least one positive test was obtained with sera collected, on average, 5 days (range, 2 to 10 days) prior to the first positive blood culture, while blood cultures were constantly negative . High and persistent mannanemias were detected in all patients during the neutropenic period . In five patients, an increased antibody response was detected when the patients recovered from aplasia . Controls consisted of 48 serum samples from 12 febrile neutropenic patients with aspergillosis (n = 4), bacteremia (n = 4), or no evidence of infection (n = 4) . A low level of mannanemia was detected in only one serum sample, and none showed significant Candida antibody titers . Our data thus confirm the value of the combined detection of mannanemia and antimannan antibodies in individuals at risk of candidemia and suggest that in neutropenic patients, an approach based on the regular monitoring of both markers could contribute to the earlier diagnosis of C . tropicalis systemic infection.

Appl Environ Microbiol, 2003 Oct, 69(10), 6179 - 88
Cloning and characterization of the xyl1 gene, encoding an NADH-preferring xylose reductase from Candida parapsilosis, and its functional expression in Candida tropicalis; Lee JK et al.; Xylose reductase (XR) is a key enzyme in D-xylose metabolism, catalyzing the reduction of D-xylose to xylitol . An NADH-preferring XR was purified to homogeneity from Candida parapsilosis KFCC-10875, and the xyl1 gene encoding a 324-amino-acid polypeptide with a molecular mass of 36,629 Da was subsequently isolated using internal amino acid sequences and 5' and 3' rapid amplification of cDNA ends . The C . parapsilosis XR showed high catalytic efficiency (kcat/Km = 1.46 s(-1) mM(-1)) for D-xylose and showed unusual coenzyme specificity, with greater catalytic efficiency with NADH (kcat/Km = 1.39 x 10(4) s(-1) mM(-1)) than with NADPH (kcat/Km = 1.27 x 10(2) s(-1) mM(-1)), unlike all other aldose reductases characterized . Studies of initial velocity and product inhibition suggest that the reaction proceeds via a sequentially ordered Bi Bi mechanism, which is typical of XRs . Candida tropicalis KFCC-10960 has been reported to have the highest xylitol production yield and rate . It has been suggested, however, that NADPH-dependent XRs, including the XR of C . tropicalis, are limited by the coenzyme availability and thus limit the production of xylitol . The C . parapsilosis xyl1 gene was placed under the control of an alcohol dehydrogenase promoter and integrated into the genome of C . tropicalis . The resulting recombinant yeast, C . tropicalis BN-1, showed higher yield and productivity (by 5 and 25%, respectively) than the wild strain and lower production of by-products, thus facilitating the purification process . The XRs partially purified from C . tropicalis BN-1 exhibited dual coenzyme specificity for both NADH and NADPH, indicating the functional expression of the C . parapsilosis xyl1 gene in C . tropicalis BN-1 . This is the first report of the cloning of an xyl1 gene encoding an NADH-preferring XR and its functional expression in C . tropicalis, a yeast currently used for industrial production of xylitol.

Appl Environ Microbiol, 2003 Oct, 69(10), 5992 - 9
Transformation of fatty acids catalyzed by cytochrome P450 monooxygenase enzymes of Candida tropicalis; Eschenfeldt WH et al.; Candida tropicalis ATCC 20336 can grow on fatty acids or alkanes as its sole source of carbon and energy, but strains blocked in beta-oxidation convert these substrates to long-chain alpha,omega-dicarboxylic acids (diacids), compounds of potential commercial value (Picataggio et al., Biotechnology 10:894-898, 1992) . The initial step in the formation of these diacids, which is thought to be rate limiting, is omega-hydroxylation by a cytochrome P450 (CYP) monooxygenase . C . tropicalis ATCC 20336 contains a family of CYP genes, and when ATCC 20336 or its derivatives are exposed to oleic acid (C(18:1)), two cytochrome P450s, CYP52A13 and CYP52A17, are consistently strongly induced (Craft et al., this issue) . To determine the relative activity of each of these enzymes and their contribution to diacid formation, both cytochrome P450s were expressed separately in insect cells in conjunction with the C . tropicalis cytochrome P450 reductase (NCP) . Microsomes prepared from these cells were analyzed for their ability to oxidize fatty acids . CYP52A13 preferentially oxidized oleic acid and other unsaturated acids to omega-hydroxy acids . CYP52A17 also oxidized oleic acid efficiently but converted shorter, saturated fatty acids such as myristic acid (C(14:0)) much more effectively . Both enzymes, in particular CYP52A17, also oxidized omega-hydroxy fatty acids, ultimately generating the alpha,omega-diacid . Consideration of these different specificities and selectivities will help determine which enzymes to amplify in strains blocked for beta-oxidation to enhance the production of dicarboxylic acids . The activity spectrum also identified other potential oxidation targets for commercial development.

Appl Environ Microbiol, 2003 Oct, 69(10), 5983 - 91
Identification and characterization of the CYP52 family of Candida tropicalis ATCC 20336, important for the conversion of fatty acids and alkanes to alpha,omega-dicarboxylic acids; Craft DL et al.; Candida tropicalis ATCC 20336 excretes alpha,omega-dicarboxylic acids as a by-product when cultured on n-alkanes or fatty acids as the carbon source . Previously, a beta-oxidation-blocked derivative of ATCC 20336 was constructed which showed a dramatic increase in the production of dicarboxylic acids . This paper describes the next steps in strain improvement, which were directed toward the isolation and characterization of genes encoding the omega-hydroxylase enzymes catalyzing the first step in the omega-oxidation pathway . Cytochrome P450 monooxygenase (CYP) and the accompanying NADPH cytochrome P450 reductase (NCP) constitute the hydroxylase complex responsible for the first and rate-limiting step of omega-oxidation of n-alkanes and fatty acids . 10 members of the alkane-inducible P450 gene family (CYP52) of C . tropicalis ATCC20336 as well as the accompanying NCP were cloned and sequenced . The 10 CYP genes represent four unique genes with their putative alleles and two unique genes for which no allelic variant was identified . Of the 10 genes, CYP52A13 and CYP52A14 showed the highest levels of mRNA induction, as determined by quantitative competitive reverse transcription-PCR during fermentation with pure oleic fatty acid (27-fold increase), pure octadecane (32-fold increase), and a mixed fatty acid feed, Emersol 267 (54-fold increase) . The allelic pair CYP52A17 and CYP52A18 was also induced under all three conditions but to a lesser extent . Moderate induction of CYP52A12 was observed . These results identify the CYP52 and NCP genes as being involved in alpha,omega-dicarboxylic acid production by C . tropicalis and provide the foundation for biocatalyst improvement.

Medicine (Baltimore), 2003 Sep, 82(5), 309 - 21
Candidemia in a tertiary care cancer center: in vitro susceptibility and its association with outcome of initial antifungal therapy; Antoniadou A et al.; Since the 1990s, changing trends have been documented in species distribution and susceptibility to bloodstream infections caused by Candida species in cancer patients . However, few data are available regarding the association between in vitro antifungal susceptibility and outcome of candidemia in this patient population . We therefore evaluated the association of in vitro antifungal susceptibility and other risk factors with failure of initial antifungal therapy in cancer patients with candidemia . Candidemia cases in cancer patients from 1998 to 2001 (n = 144) were analyzed retrospectively along with their in vitro susceptibility to amphotericin B, fluconazole, and itraconazole (National Committee for Clinical and Laboratory Standards M27-A method) . Patients were evaluable for outcome analysis if they received continuous unchanged therapy with either fluconazole or amphotericin B for >/=5 days . We excluded cases of mixed candidemia . In vitro susceptibility testing data of the first Candida bloodstream isolate were analyzed . Appropriate therapy was defined as that using an active in vitro antifungal for >/=5 days . For fluconazole susceptible-dose dependent Candida species, we defined appropriate therapy as a fluconazole dose of >/=600 mg/day . The Candida species distribution was 30% Candida albicans, 24% Candida glabrata, 23% Candida parapsilosis, 10% Candida krusei, 9% Candida tropicalis, and 3% other . Overall, amphotericin B was the most active agent in vitro, with only 3% of the isolates exhibiting resistance to it (>1 mg/L) . Dose-dependent susceptibility to fluconazole and itraconazole was seen in 13% and 21% of the isolates, respectively, while resistance to fluconazole and itraconazole was seen in 13% and 26%, respectively.Eighty patients were evaluable for outcome analysis . In multivariate analysis, the following factors emerged as independent predictors of failure of initial antifungal therapy: leukemia (p = 0.01), bone marrow transplantation (p = 0.006), and intensive care unit stay at onset of infection (p = 0.02) . Inappropriate antifungal therapy, as defined by daily dose and in vitro susceptibility, was not shown consistently to be a significant factor (it was significant in multivariate analysis, p = 0.04, but not in univariate analysis), indicating the complexity of the variables that influence the response to antifungal treatment in cancer patients with candidemia.

Transplant Proc, 2003 Sep, 35(6), 2298 - 303
Epidemiology and susceptibility to antifungal agents of fungi isolated from clinical specimens from patients hospitalized in the Department of General and Liver Surgery of the Medical University of Warsaw; Swoboda-Kopec E et al.; The aim of this study was to analyze the type and antibiotic susceptibility of fungi isolated from clinical specimens obtained from patients hospitalized in the Department of General, Transplantation and Liver Surgery of the Medical University of Warsaw between 2000 to 2002 . Among the 326 clinical samples found to be positive on mycological culture, 356 strains were cultured . The most common isolates were yeastlike fungi of the genus Candida 334 (93.8%), while others included 33 other types (6.2%) . The most commonly isolated species were Candida albicans, 194 strains (54.5%); Candida glabrata, 68 (19.1%); Candida krusei, 20 (5.6%); Candida inconspicua, 20 (5.6%); Candida tropicalis, 17 (4.8%); and Candida parapsilosis, 6 (1.7%) . Upon testing for susceptibility to antifungal agents, all strains were susceptible to amphotericin B, while 43.8% of strains showed intermediate susceptibility to fluconazole and 25.3%, to itraconazole . Control of fungal infections in transplant and in immunocompromised patients is hindered by the low percentage of strains susceptible to commonly used antifungal agents, particularly of the triazole group.

Chemotherapy, 2003 Sep, 49(5), 243 - 7
Brief exposure to antimycotics reduces the extracellular phospholipase activity of Candida albicans and Candida tropicalis; Anil S et al.; BACKGROUND: Although the phospholipase activity is considered a potential virulence determinant of the pathogenic Candida species, the effect of antimycotics on this attribute is not known . Hence we evaluated the phospholipase activity in 10 isolates each of Candida albicans and Candida tropicalis, after their exposure to antifungals . METHODS: The impact of antimycotics on phospholipase activity was also assessed after exposure of the isolates to sub-minimum inhibitory concentrations of nystatin, amphotericin B and fluconazole . RESULTS: All Candida isolates investigated exhibited phospholipase activity (Pz) . In general C . ALBICANS showed relatively higher P(z) activity than C . tropicalis , and exposure of the isolates to antimycotics led to a significant (p < 0.05) reduction in the phospholipase activity . Nystatin and amphotericin B, but not fluconazole, significantly reduced the phospholipase activity of both Candida species . CONCLUSION: These observations, while confirming the higher virulence of C . albicans relative to C . tropicalis, demonstrate for the first time the effect of antifungal agents on extracellular phospholipases of these common opportunistic pathogens .

Eur J Clin Microbiol Infect Dis, 2003 Oct, 22(10), 603 - 7 Epub 2003 Sep 12.
High-dose liposomal amphotericin B in the therapy of systemic candidiasis in neonates; Juster-Reicher A et al.; High-dose (5-7 mg/kg/day) liposomal amphotericin B was evaluated prospectively during the period 1995-2001 in 41 episodes of systemic candidiasis occurring in 37 neonates (36 of the 37 were premature infants with very low birth weights) . Median age at the onset of systemic candidiasis was 17 days . Candida spp . were isolated from blood in all patients and from urine, skin abscesses and peritoneal fluid in 6, 5 and 1 neonates, respectively . Candidiasis was due to Candida parapsilosis in 17 cases, Candida albicans in 15 cases, Candida tropicalis in 5 cases, Candida guilliermondii in 2 cases, Candida glabrata in 2 cases and an unidentified Candida sp . in 1 case . Twenty-eight, five and eight infants received 7, 6-6.5 and 5 mg/kg/day, respectively . Median duration of therapy was 18 days; median cumulative dose was 94 mg/kg . Fungal eradication was achieved in 39 of 41 (95%) episodes; median duration of therapy until fungal eradication was 8.7+/-4.5 days . Fungal eradication was achieved after 10.9+/-4.8 days in patients who had received previous antifungal therapy compared to 8.2+/-4.3 days in those treated with liposomal amphotericin B as first-line therapy . One patient died due to systemic candidiasis on day 12 of therapy . High-dose liposomal amphotericin B was effective and safe in the treatment of neonatal candidiasis . Fungal eradication was more rapid in patients treated early with high doses and in patients who received high-dose liposomal amphotericin B as first-line therapy.

Clin Diagn Lab Immunol, 2003 Sep, 10(5), 835 - 48
Competitive binding inhibition enzyme-linked immunosorbent assay that uses the secreted aspartyl proteinase of Candida albicans as an antigenic marker for diagnosis of disseminated candidiasis; Morrison CJ et al.; The secreted aspartyl proteinases (Saps) of Candida albicans have been implicated as virulence factors associated with adherence and tissue invasion . The potential use of proteinases as markers of invasive candidiasis led us to develop a competitive binding inhibition enzyme-linked immunosorbent assay (ELISA) to detect Sap in clinical specimens . Daily serum and urine specimens were collected from rabbits that had been immunosuppressed with cyclophosphamide and cortisone acetate and infected intravenously with 10(7) C . albicans blastoconidia . Disseminated infection was confirmed by organ culture and histopathology . Although ELISA inhibition was observed when serum specimens from these rabbits were used, more significant inhibition, which correlated with disease progression, occurred when urine specimens were used . Urine collected as early as 1 day after infection resulted in significant ELISA inhibition (mean inhibition +/- standard error {SE} compared with preinfection control urine, 15.7% +/- 2.7% {P < 0.01}), and inhibition increased on days 2 through 5 (29.4% +/- 4.8% to 44.5% +/- 3.5% {P < 0.001}) . Urine specimens from immunosuppressed rabbits infected intravenously with Candida tropicalis, Candida parapsilosis, Candida krusei, Cryptococcus neoformans, Aspergillus fumigatus, or Staphylococcus aureus were negative in the assay despite culture-proven dissemination . Nonimmunosuppressed rabbits receiving oral tetracycline and gentamicin treatment were given 2 x 10(8) C . albicans blastoconidia orally or intraurethrally to establish colonization of the gastrointestinal tract or bladder, respectively, without systemic dissemination; urine specimens from these rabbits also gave negative ELISA results . Dissemination to the kidney and spleen occurred in one rabbit challenged by intragastric inoculation, and urine from this rabbit demonstrated significant inhibition in the ELISA (mean inhibition +/- SE by day 3 after infection, 32.9% +/- 2.7% {P < 0.001}) . The overall test sensitivity was 83%, the specificity was 92%, the positive predictive value was 84%, the negative predictive value was 91%, and the efficiency was 89% (166 urine samples from 33 rabbits tested) . The specificity, positive predictive value, and efficiency could be increased to 97, 95, and 92%, respectively, if at least two positive test results were required for a true positive designation . The ELISA was sensitive and specific for the detection of Sap in urine specimens from rabbits with disseminated C . albicans infection, discriminated between colonization and invasive disease, reflected disease progression and severity, and has the potential to be a noninvasive means to diagnose disseminated candidiasis.

Arch Pharm (Weinheim), 2003 Aug, 336(6-7), 322 - 35
Antimycobacterial and antifungal isosters of salicylamides; Waisser K et al.; A set of 40 derivatives of 3-hydroxypicolinic acid and 2-sulfanylbenzoic acid, isosteric to salicylanilides was synthesized . The compounds were evaluated for in vitro activity against Mycobacterium tuberculosis, Mycobacterium kansasii and Mycobacterium avium, Candida albicans, Candida tropicalis, Candida krusei, Candida glabrata, Trichosporon beigelii, Aspergillus fumigatus, Absidia corymbifera, Trichophyton mentagrophytes and Microsporum gypseum . Structure-activity relationships of antimycobacterial activity and antifungal activity against T . mentagrophytes and M . gypseum were analyzed by the Free-Wilson method . An increase in antimycobacterial activity was observed only for the sulfanylbenzoic acid derivatives, especially those with the benzyl moiety . The antifungal activity was not significant.

J Antimicrob Chemother, 2003 Oct, 52(4), 679 - 82 Epub 2003 Sep 01.
The European Confederation of Medical Mycology (ECMM) survey of candidaemia in Italy: antifungal susceptibility patterns of 261 non-albicans Candida isolates from blood; Tortorano AM et al.; OBJECTIVES: To analyse the in vitro antifungal susceptibility of 261 non-albicans Candida bloodstream strains isolated during the European Confederation of Medical Mycology survey of candidaemia performed in Lombardia, Italy (September 1997-December 1999) . METHODS: In vitro susceptibility to flucytosine, fluconazole, itraconazole, posaconazole and voriconazole was determined using the broth microdilution method described in the NCCLS M27-A guidelines . Etest strips were used to assess susceptibility to amphotericin B . In vitro findings were correlated with the patient's underlying condition and previous antifungal treatment . RESULTS: MICs (mg/L) at which 90% of the strains were inhibited were, respectively, 2 for flucytosine, 8 for fluconazole, 0.5 for itraconazole, 0.25 for voriconazole and 0.25 for posaconazole . Amphotericin B MIC endpoints were <0.50 mg/L in all the isolates tested . Flucytosine resistance was detected in 19 isolates (7%), mainly among Candida tropicalis strains (30%) . Innate or secondary fluconazole resistance was detected in 13 strains (5%) . Among the 13 patients with fluconazole-resistant Candida bloodstream infection, three were HIV positive, including one treated with fluconazole for oral candidosis; the four who were HIV negative had received the azole during the 2 weeks preceding the candidaemia . Cross-resistance among fluconazole and other azoles was a rare event . CONCLUSIONS: Resistance is still uncommon in non-albicans Candida species recovered from blood cultures . However, in fungaemias caused by C . tropicalis, Candida glabrata and Candida krusei, there is a high prevalence of resistance to fluconazole and flucytosine . Fluconazole resistance should be suspected in patients treated previously with azoles, mainly those with advanced HIV infection.

Mycoses, 2003 Sep, 46(8), 287 - 92
An outbreak of candidemia due to Candida tropicalis in a neonatal intensive care unit; Chowdhary A et al.; An outbreak of candidemia due to Candida tropicalis involving 16 neonates (gestational age 28-36 weeks) is reported . All infants had received hyperalimentation and at least one course of antibiotics . The commonest clinical manifestations included episodes of acute respiratory distress and lack of response to antibacterial antibiotic therapy . Candida tropicalis was recovered from blood in all the 16 infants and urine cultures were positive in 14 infants . Environmental sampling yielded C . tropicalis from one each of the blankets and mattresses used for neonates . Four of five urinary tract isolates and both environmental isolates genotyped by arbitrarily primed-PCR with several random primers were shown to belong to the same genotype.

Cell Biochem Funct, 2003 Sep, 21(3), 269 - 74
A combination of alpha-tocopherol, vitamin C and N-acetyl cysteine increases unsaturated fatty acid levels in hydrogen peroxide-induced Candida tropicalis (ATCC 13803); Yilmaz O et al.; This research was aimed at evaluating the antioxidant effects of combinations of alpha lipoic acid (LA), vitamin C (VC), N-acetyl cysteine (NAC) and alpha-tocopherol (TOC) on lipid level and fatty acid composition of C . tropicalis (ATCC 13803) against hydrogen peroxide toxicity . According to the experimental results, the cell density of C . tropicalis increased significantly in NAC+LA+H2O2, NAC+TOC+ H2O2 and NAC+VC+H2O2 groups (p<0.001) at the end of 48 and 72 h incubation times . The total lipid level in H2O2 and H2O2 + antioxidant-supplemented groups was lower than that of the control group . In the fatty acid composition of C . tropicalis, the palmitic acid level was raised in the NAC group (p<0.05), whereas its level was reduced in the other supplemented groups . While the oleic acid level increased in NAC+TOC+H2O2 and NAC+VC+H2O2 (p<0.001) groups, its level slightly decreased in the H2O2 group . The linolenic acid level was low in all the supplemented groups, but linoleic acid and total mono-unsaturated fatty acid (MUFA) levels were high in these groups compared with the control group . Total polyunsaturated fatty acid level (PUFA) decreased in NAC and H2O2 groups (p<0.01), but its level increased in NAC+LA+H2O2 and NAC+TOC+H2O2 groups (respectively, p<0.01, p<0.001) . Total saturated fatty acid level decreased significantly in NAC+TOC+H2O2, NAC+H2O2 and NAC+VC+H2O2 (p<0.001) groups (p<0.01), whereas total unsaturated fatty acid level increased in NAC, NAC+H2O2, NAC+LA+H2O2, NAC+TOC+H2O2 and NAC+VC+H2O2 groups . In conclusion, our data showed that the levels of total unsaturated fatty acid, MUFA and PUFA were raised with the combinations of NAC and TOC, LA and VC in C . tropicalis cells subjected to hydrogen peroxide toxicity .

Int J Food Microbiol, 2003 Sep 1, 86(1-2), 189 - 200
CHROMagar Candida medium as a practical tool for the differentiation and presumptive identification of yeast species isolated from salads; Tornai-Lehoczki J et al.; CHROMagar Candida medium was used to study the diversity of yeast biota of salad samples, and to presumptively identify the isolates . This medium was originally developed for the selective isolation and presumptive identification of some clinically important yeast species such as Candida albicans, Candida tropicalis, Candida krusei, and Candida glabrata on the basis of differences in colour and surface of colonies . Ninety three yeast strains representing 33 species from the culture collection and 39 fresh isolates from different mayonnaise-based mixed salads showed a wide range of hue of colony colours ranging from white to yellow, orange, red, pink, purple, blue, green, etc., as well as different morphological appearances on the CHROMagar Candida medium . Therefore, CHROMagar Candida medium facilitates the detection of mixtures of yeast species from different samples on a single isolation plate and this medium can be a practical method for the differentiation and rapid presumptive identification of many yeast species occurring frequently in different kind of foods.

Int J Food Microbiol, 2003 Sep 1, 86(1-2), 87 - 99
The microbial ecology of cocoa bean fermentations in Indonesia; Ardhana MM et al.; Cocoa beans are the principal raw material of chocolate manufacture . The beans are subject to a microbial fermentation as the first stage in chocolate production . The microbial ecology of bean fermentation (Forastero and Trinitario cultivars) was investigated at three commercial fermentaries in East Java, Indonesia by determining the populations of individual species at 12-h intervals throughout the process . The first 2-3 days of fermentation were characterised by the successional growth of various species of filamentous fungi, yeasts, lactic acid bacteria and acetic acid bacteria . The principal species found were Penicillium citrinum, an unidentified basidiomycete, Kloeckera apis, Saccharomyces cerevisiae, Candida tropicalis, Lactobacillus cellobiosus, Lactobacillus plantarum and Acetobacter pasteurianus . The later stages of fermentation were dominated by the presence of Bacillus species, mostly, Bacillus pumilus and Bacillus licheniformis . Glucose, fructose, sucrose and citric acid of the bean pulp were utilised during fermentation, with the production of ethanol, acetic acid and lactic acid that diffused into the beans . The filamentous fungi were notable for their production of polygalacturonase activity and probably contributed to the degradation of bean pulp.

J Biol Chem, 2003 Oct 17, 278(42), 41213 - 20 Epub 2003 Jul 30.
Candida tropicalis expresses two mitochondrial 2-enoyl thioester reductases that are able to form both homodimers and heterodimers; Torkko JM et al.; Here we report on the cloning of a Candida tropicalis gene, ETR2, that is closely related to ETR1 . Both genes encode enzymatically active 2-enoyl thioester reductases involved in mitochondrial synthesis of fatty acids (fatty acid synthesis type II) and respiratory competence . The 5'- and 3'-flanking (coding) regions of ETR2 and ETR1 are about 90% (97%) identical, indicating that the genes have evolved via gene duplication . The gene products differ in three amino acid residues: Ile67 (Val), Ala92 (Thr), and Lys251 (Arg) in Etr2p (Etr1p) . Quantitative PCR analysis and reverse transcriptase-PCR indicated that both genes were expressed about equally in fermenting and ETR1 predominantly respiring yeast cells . Like the situation with ETR1, expression of ETR2 in respiration-deficient Saccharomyces cerevisiae mutant cells devoid of Ybr026p/Etr1p was able to restore growth on glycerol . Triclosan that is used as an antibacterial agent against fatty acid synthesis type II 2-enoyl thioester reductases inhibited growth of FabI overexpressing mutant yeast cells but was not able to inhibit respiratory growth of the ETR2- or ETR1-complemented mutant yeast cells . Resolving of crystal structures obtained via Etr2p and Etr1p co-crystallization indicated that all possible dimer variants occur in the same asymmetric unit, suggesting that similar dimer formation also takes place in vivo.

Malays J Pathol, 2002 Dec, 24(2), 83 - 9
Risk assessment and microbiological profile of infections in paediatric cancer patients with febrile neutropenia; Latiff Z et al.; Febrile neutropenia is a common and potentially fatal problem encountered in cancer patients undergoing chemotherapy . We carried out an observational study to evaluate the possible risk factors of developing fever amongst neutropenic children with an underlying malignancy . We also looked at the microbiological profile of causative pathogens in patients with febrile neutropenia . During a study period of 1 year, a total of 90 neutropenic episodes were recorded amongst 57 patients who were on treatment and follow-up during the study period . Multivariate analysis showed that factors such as chemotherapy status, underlying disease, existing central venous catheters, presenting white blood cell counts at chemotherapy, use of steroid therapy or hospitalisation at the onset of neutropenia, were not significant risk factors for developing fever during neutropenic episodes . Although the presence of a central venous catheter was associated with a higher risk of developing fever, it did not reach statistical significance (p=0.11) . Of the 90 neutropenic episodes, 59 (65.6%) developed fever and 25 of these had positive blood cultures . The causative organisms include gram-negative bacteria (64%), gram positive bacteria (16%) and fungus (20%) . Of the gram-negative organisms, Klebsiella spp . predominated (28%) with the extended spectrum beta-lactamase producing strain forming the majority (16%) . Amongst those with fungaemia, Candida spp . and Candida tropicalis formed the majority (8% each) of the isolates.

Eur J Clin Microbiol Infect Dis, 2003 Aug, 22(8), 470 - 4 Epub 2003 Jul 23.
Global distribution and outcomes for Candida species causing invasive candidiasis: results from an international randomized double-blind study of caspofungin versus amphotericin B for the treatment of invasive candidiasis; Colombo AL et al.; In a randomized study, caspofungin was compared with amphotericin B for the treatment of invasive candidiasis in a total of 239 adults from 56 sites in 20 countries . This study provided a unique opportunity to assess the frequency and outcome of invasive candidiasis caused by different Candida species worldwide, and the results are presented here . Efficacy was primarily assessed at the end of intravenous therapy using a modified intent-to-treat (MITT) analysis . This analysis was performed on 224 of the 239 patients enrolled in the study . Attempts were made to collect baseline Candida isolates from all patients for species identification at a central laboratory . Yeasts were identified to the species level using two commercial systems and microscopic examination . Viable baseline isolates were recovered from 210 of the 224 (94%) patients included in the MITT analysis . Candida albicans was the most frequently isolated species in all regions and was responsible for 45% of cases overall . Nevertheless, the majority of cases of infection were caused by non- albicans Candida species . In the USA and Canada, Candida glabrata was the second most commonly isolated pathogen (18%) . In contrast, Candida parapsilosis and Candida tropicalis accounted for 55% of cases in Latin America . Outcomes were comparable for patients treated with caspofungin (74% overall; 64% and 80% for infections due to Candida albicans and non- albicans species) and amphotericin B (62% overall; 58% and 68% for infections due to Candida albicans and non- albicans species), and were generally similar across continents . The distribution of Candida species isolated from patients enrolled in a clinical trial may not be representative of pathogens causing invasive candidiasis in the general population . Nevertheless, our findings may affect the regional choice of empirical antifungal therapy for seriously ill patients with suspected or documented invasive candidiasis since different Candida species have varying susceptibility to conventional antifungal drugs.

Biotechnol Lett, 2003 May, 25(9), 709 - 13
Decolorization of synthetic dyes and production of manganese-dependent peroxidase by new fungal isolates; Yang Q et al.; Two yeasts, Debaryomyces polymorphus, Candida tropicalis, and two filamentous fungi, Umbelopsis isabellina, Penicillium geastrivorus, could completely decolorize 100 mg Reactive Black 5 (RB 5) l-1 within 16-48 h . Manganese-dependent peroxidase (MnP) activities between 60 and 424 U l-1 were detected in culture supernatants of three of these organisms indicating the color removal by enzymatic biodegradation but with P . geastrivorus there was no ligninolytic enzyme activity in its culture and the decolorization was mainly due to biosorption to mycelium . Extensive decolorization by D . polymorphus (69-94%) and C . tropicalis (30-97%) was obtained with five other azo dyes and one anthraquinone dye . Except for Reactive Brilliant Blue KNR and Reactive Yellow M-3R, the four azo dyes, Reactive Red M-3BE, Procion Scharlach H-E3G, Procion Marine H-EXL and Reactive Brilliant Red K-2BP, induced D . polymorphus to produce MnP (105-587 U l-1) . However, MnP activities of 198-329 U l-1 were only detected in the culture of C . tropicalis containing Reactive Red M-3BE and Reactive Brilliant Red K-2BP, respectively.

Clin Infect Dis, 2003 Jul 15, 37(2), 208 - 13 Epub 2003 Jul 09.
Fungal infections in patients with severe acute pancreatitis and the use of prophylactic therapy; De Waele JJ et al.; Data from an 8-year period for 46 patients with severe acute pancreatitis and infected pancreatic necrosis were analyzed to determine the incidence of fungal infection, to identify risk factors for the development of fungal infection, and to assess the use of early fluconazole treatment . Intraabdominal fungal infection was found in 17 (37%) of 46 patients . Candida albicans was isolated most frequently (15 patients); Candida tropicalis and Candida krusei were found in 1 patient each . Characteristics of patients with fungal infection were not different from patients without fungal infection . The difference in mortality was not statistically significant between patients with fungal infection and patients without fungal infection . Early antifungal therapy (prophylactic or preemptive antifungal therapy) was administered to 18 patients, and only 3 of them developed fungal infection . In this cohort of critically ill patients, no risk factors for fungal infection could be demonstrated, and mortality among patients who received early antifungal therapy was not different . Early treatment with fluconazole seems to prevent fungal infection in these high-risk patients.

Bioorg Med Chem Lett, 2003 Aug 4, 13(15), 2601 - 5
1,2,4-Triazolo mercapto and aminonitriles as potent antifungal agents; Collin X et al.; A series of 3-mercapto-1,2,4-triazoles mono or disubstituted at 2-, 3- or 4-positions were synthesized and evaluated as antifungal agents . Many of these derivatives exhibit high activity against Candida albicans and Candida tropicalis.

Yeast, 2003 Jul 15, 20(9), 803 - 11
ALG2, the Hansenula polymorpha isocitrate lyase gene; Berardi E et al.; To set the basis for molecular and cellular studies of the glyoxylate cycle in methylotrophic yeasts, we isolated and characterized ALG2, the Hansenula polymorpha isocitrate lyase gene . Complementation work and sequence analysis revealed an ORF of 1458 nucleotides, encoding a 486 amino acid protein with a predicted molecular mass of 54.9 kDa . This protein is shorter than the Saccharomyces cerevisiae and Candida tropicalis ICLs, lacks a PST1 signal and possesses a PTS2-like signal . The transcriptional regulation of ALG2 mRNA levels by carbon source is mainly achieved by glucose repression-derepression, whereas ethanol induction plays only a minor role . We present evidence indicating that, in H . polymorpha, neither isocitrate lyase activity nor the ALG2 gene product are necessary for C(1)-peroxisome degradation triggered by ethanol . Therefore, the involvement of glyoxylate in degradation, as described by Kulachkovsky et al . (1997) for Pichia methanolica, does not necessarily apply to all methylotrophic yeasts . The relevant nucleotide sequence has been deposited at GenBank (Accession No . AF373067.1) .

Scand J Infect Dis, 2003, 35(4), 288 - 91
An unusual cause of vertebral osteomyelitis: Candida species; Garbino J et al.; Candida species rarely cause spondylodiscitis . During 3 y, 3 cases of vertebral osteomyelitis due to Candida spp . (Candida albicans and Candida tropicalis) were diagnosed, 2 of which were associated with a spinal epidural abscess.

Cancer, 2003 Jul 1, 98(1), 24 - 30
Candidemia in women with breast carcinoma treated with high-dose chemotherapy and autologous bone marrow transplantation; Gottfredsson M et al.; BACKGROUND: Invasive fungal infections, including candidemia, pose a major threat to patients with impaired immune defenses, including bone marrow transplantation (BMT) recipients . During 1992-1997, 845 women with multiple lymph node positive or metastatic breast carcinoma underwent high-dose chemotherapy (HDC) and subsequent autologous BMT at Duke University Medical Center . No systemic antifungal prophylaxis was administered . The purpose of the current study was to evaluate the risk and long-term outcome of candidemia in this patient population . METHODS: Clinical data were collected on patients with candidemia, and a group of age-matched control patients were identified who underwent HDC and BMT for breast carcinoma in the same time period . The difference in crude mortality between these two groups was used to calculate the attributable mortality of candidemia . The genetic relatedness of the fungal blood stream isolates was investigated by DNA fingerprinting . Antifungal susceptibility testing was performed using serial microdilution . RESULTS: Twenty-nine of 845 women developed candidemia (3.4%) . The crude mortality of women with candidemia was 35% at 90 days after transplantation but 11% among women in the matched control group who were without infection (P = 0.01), for an attributable mortality rate of 24% . The most common pathogen was Candida tropicalis (50%), followed by Candida albicans (23%) . The mortality was highest for women who were infected with C . albicans, followed by C . tropicalis, and other Candida species (P = 0.037) . DNA fingerprinting of the yeasts revealed genetic heterogeneity in all species . However, 9 of 15 C . tropicalis isolates had identical DNA fingerprint profiles, suggesting spread of this genotype from a common source . All yeast isolates were susceptible to amphotericin B, and 20 of 30 isolates were susceptible to <or= 8 microg/mL of fluconazole . CONCLUSIONS: Candidemia was relatively infrequent after HDC and autologous BMT in women with for multiple lymph node positive or metastatic breast carcinoma . This was true even in the absence of systemic antifungal prophylaxis . The mortality attributable to candidemia in this patient population was 24% and was higher among patients who were infected by C . albicans compared with patients who were infected by other Candida, non-albicans species .

Acta Crystallogr D Biol Crystallogr, 2003 Jul, 59(Pt 7), 1302 - 5 Epub 2003 Jun 27.
Crystallization and preliminary crystallographic data of 2-enoyl-CoA hydratase 2 domain of Candida tropicalis peroxisomal multifunctional enzyme type 2; Koski MK et al.; In yeast, the second and the third reaction of the fatty-acid beta-oxidation spiral are catalysed by peroxisomal multifunctional enzyme type 2 (Mfe2p/Fox2p) . This protein has two (3R)-hydroxyacyl-CoA dehydrogenase domains and a C-terminal 2-enoyl-CoA hydratase 2 domain . Here, the purification, crystallization and X-ray diffraction analysis of the hydratase 2 domain {CtMfe2p(dh(a+b)Delta)} from Candida tropicalis Mfe2p is reported . CtMfe2p(dh(a+b)Delta) was overexpressed as an enzymatically active recombinant protein and crystallized by the hanging-drop vapour-diffusion method . The crystals belong to space group C2, with unit-cell parameters a = 178.57, b = 60.46, c = 130.85 A, beta = 94.48 degrees . Selenomethionine-labelled protein was used for a multi-wavelength anomalous dispersion (MAD) experiment . A three-wavelength data set suitable for MAD phasing was collected to 2.25 A resolution using synchrotron radiation.

FEMS Microbiol Lett, 2003 Jun 27, 223(2), 215 - 9
Biodegradation of polyphenols with immobilized Candida tropicalis under metabolic induction; Ettayebi K et al.; During olive oil production, large quantities of olive mill wastewater (OMW) are produced . This wastewater material, containing a high level of phenolic compounds, poses a serious environmental problem in almost all Mediterranean countries . Candida tropicalis YMEC14 was used as an extremophile strain to design an aerobic biotreatment process to detoxify OMW and reduce its polluting organic load . The process was enhanced by directing yeast metabolism towards biodegradation pathways using hexadecane as co-metabolite and by immobilizing yeast cells in calcium alginate beads . Under immobilization conditions, C . tropicalis YMEC14 grown at 40 degrees C in OMW supplemented with hexadecane resulted in 69.7%, 69.2% and 55.3% reduction of chemical oxygen demand, monophenols and polyphenols, respectively, after a 24-h fermentation cycle.

Antimicrob Agents Chemother, 2003 Jul, 47(7), 2208 - 16
Preclinical assessment of the efficacy of mycograb, a human recombinant antibody against fungal HSP90; Matthews RC et al.; Mycograb (NeuTec Pharma plc) is a human genetically recombinant antibody against fungal heat shock protein 90 (HSP90) . Antibody to HSP90 is closely associated with recovery in patients with invasive candidiasis who are receiving amphotericin B (AMB) . Using in vitro assays developed for efficacy assessment of chemotherapeutic antifungal drugs, Mycograb showed activity against a wide range of yeast species (MICs against Candida albicans {fluconazole {FLC}-sensitive and FLC-resistant strains}, Candida krusei, Candida tropicalis, Candida glabrata, and Candida parapsilosis, 128 to 256 microg/ml) . Mycograb (4 or 8 microg/ml) showed synergy with AMB, the fractional inhibitory index being 0.09 to 0.31 . Synergy was not evident with FLC, except for FLC-sensitive C . albicans . Murine kinetics showed that Mycograb at 2 mg/kg produced a maximum concentration of drug in serum of 4.7 microg/ml, a half-life at alpha phase of 3.75 min, a half-life at beta phase of 2.34 h, and an area under the concentration-time curve from 0 to t h of 155 microg . min/ml . Mycograb (2 mg/kg) alone produced significant improvement in murine candidiasis caused by each species: (i) . a reduction (Scheffe's test, P < 0.05) in the mean organ colony count for the FLC-resistant strain of C . albicans (kidney, liver, and spleen), C . krusei (liver and spleen), C . glabrata (liver and spleen), C . tropicalis (kidney), and C . parapsilosis (kidney, liver, and spleen) and (ii) . a statistically significant increase in the number of negative biopsy specimens (Fisher's exact test, P < 0.05) for C . glabrata (kidney), C . tropicalis (liver and spleen), and C . parapsilosis (liver) . AMB (0.6 mg/kg) alone cleared the C . tropicalis infection but failed to clear infections caused by C . albicans, C . krusei, C . glabrata, or C . parapsilosis . Synergy with AMB, defined as an increase (Fisher's exact test, P < 0.05) in the number of negative biopsy specimens compared with those obtained using AMB alone, occurred with the FLC-resistant strain of C . albicans (kidney), C . krusei (spleen), C . glabrata (spleen), and C . parapsilosis (liver and spleen) . Only by combining Mycograb with AMB was complete resolution of infection achieved for C . albicans, C . krusei, and C . glabrata.

Arch Microbiol, 2003 Jul, 180(1), 76 - 80 Epub 2003 Jun 07.
Distribution of antigenic oligomannosyl side chains in the cell wall mannans of several strains of Candida tropicalis; Kobayashi H et al.; In order to clarify the distribution of antigenic oligomannosyl side chains in the cell wall mannans of the pathogenic yeast Candida tropicalis, the chemical structure of mannans isolated from four C . tropicalis strains was investigated using nuclear magnetic resonance, two-dimensional homonuclear Hartmann-Hahn (2D-HOHAHA) spectroscopy . Two-dimensional maps of the 2D-HOHAHA clearly showed the distribution of oligomannosyl side chains in the mannans . The linear side chain Manalpha1-3Manalpha1-(2Manalpha1-)(n)2Man {n> or =2} is present in the mannans from C . tropicalis IFO 0589 and IFO 1400, but not in the mannans from IFO 0199 and IFO 1647 . The mannan of IFO 0589 is the only mannan with the branched side chains, Manalpha1-3{Manalpha1-6}Manalpha1-(2Manalpha1-)(n)2Man and Manalpha1-2Manalpha1-3{Manalpha1-6}Manalpha1-(2Manalpha1-)(n)2Man {n> or =2} . However, this mannan lacked the phosphate group and the beta-1,2-linked oligomannosyl side chain which are features of this group . The mannans of the C . tropicalis strains IFO 0589 and IFO 1400 possessed the side chains containing an alpha-1,3-linked mannose residue previously observed in Candida albicans.

J Clin Microbiol, 2003 Jun, 41(6), 2629 - 32
Direct isolation of Candida spp . from blood cultures on the chromogenic medium CHROMagar Candida; Horvath LL et al.; CHROMagar Candida is a selective and differential chromogenic medium that has been shown to be useful for identification of Candida albicans, Candida krusei, Candida tropicalis, and perhaps Candida glabrata . Colony morphology and color have been well defined when CHROMagar Candida has been used to isolate yeast directly from clinical specimens, including stool, urine, respiratory, vaginal, oropharyngeal, and esophageal sources . Direct isolation of yeast on CHROMagar Candida from blood cultures has not been evaluated . We evaluated whether the color and colony characteristics produced by Candida spp . on CHROMagar Candida were altered when yeasts were isolated directly from blood cultures . Fifty clinical isolates of Candida were inoculated into aerobic and anaerobic blood culture bottles and incubated at 35 degrees C in an automated blood culture system . When growth was detected, an aliquot was removed and plated onto CHROMagar Candida . As a control, CHROMagar Candida plates were inoculated with the same isolate of yeast grown on Sabouraud dextrose agar simultaneously . No significant difference was detected in color or colony morphology between the blood and control isolates in any of the tested organisms . All C . albicans (n = 12), C . tropicalis (n = 12), C . glabrata (n = 9), and C . krusei (n = 5) isolates exhibited the expected species-specific colony characteristics and color, whether isolated directly from blood or from control cultures . CHROMagar Candida can be reliably used for direct isolation of yeast from blood cultures . Direct isolation could allow mycology laboratories to more rapidly identify Candida spp., enable clinicians to more quickly make antifungal agent selections, and potentially decrease patient morbidity and mortality.

Biochem Biophys Res Commun, 2003 Jun 20, 306(1), 129 - 33
Selectivity of 4,5,6,7-tetrabromobenzimidazole as an ATP-competitive potent inhibitor of protein kinase CK2 from various sources; Zien P et al.; Like the previously reported 4,5,6,7-tetrabromobenzotriazole (TBBt), the structurally related 4,5,6,7-tetrabromobenzimidazole (TBBz) is a selective ATP-competitive inhibitor of protein kinase CK2 from such divergent sources as yeast, rat liver, Neurospora crassa and Candida tropicalis, with K(i) values in the range 0.5-1 microM . It is virtually inactive vs . PKA, PKC, and a very weak inhibitor of protein kinase CK1 . The corresponding tetrachlorobenzimidazole (TCBz) is a much weaker inhibitor of CK2, like tetrachlorobenzotriazole (TCBt) relative to TBBt . Bearing in mind the similarity of the van der Waals radii of Br (1.95 A) and CH(3) (2.0 A), the corresponding much less hydrophobic 4,5,6,7-tetramethylbenzotriazole (TMeBt) was prepared and found to be a very weak inhibitor of CK2, as well as of CK1 . An unexpected, and significant, difference between TBBt and TBBz are their inhibitory activities vs . the yeast protein kinase PK60S, which phosphorylates, both in vitro and in intact yeast cells, three of the five pp13 kDa ribosomal surface acidic proteins in yeast cells . TBBt was previously noted to be a more effective inhibitor of PK60S than of yeast CK2; by contrast, TBBz is a relatively feeble inhibitor of PK60S, hence more selective than TBBt vs . CK2 in yeast cells . TMeBt was virtually inactive vs PK60S . Like TBBt, TBBz is an additional lead compound for development of more potent inhibitors of CK2.

J Antimicrob Chemother, 2003 Jul, 52(1), 71 - 7 Epub 2003 May 29.
Stable susceptibility of Candida blood isolates to fluconazole despite increasing use during the past 10 years; Chen YC et al.; The prevalence of drug-resistant bacterial pathogens is very high in Taiwan . Accordingly, there was great concern that the introduction of fluconazole would result in rapid emergence of drug-resistant yeasts . Thus, we recommended in 1991 that fluconazole be used for treatment only . To explore the impact of this policy fluconazole susceptibility of Candida species blood culture isolates and outcome of patients with nosocomial candidaemia were monitored prospectively at National Taiwan University Hospital during 1994-2000 . The MICs of fluconazole were determined by the disc diffusion method . There were 1095 episodes of nosocomial candidaemia during 1994-2000 . Candida albicans was the most common species (50.4%), followed by Candida tropicalis (20.5%), Candida parapsilosis (14.2%) and Candida glabrata (12.0%) . There were 0-2 isolates of Candida krusei per year . The incidence of nosocomial candidaemia and the proportion of C . glabrata peaked in 1996 and decreased thereafter . Fluconazole susceptibility was determined for 552 Candida blood isolates . Only 0.7% of blood isolates were resistant to fluconazole . Fluconazole susceptibility was 94.0% in 1994-1995 and 97.9% in 1999-2000 (P = 0.06) . Attributable mortality for patients with nosocomial candidaemia was 43.2% in 1994-1995 and was 25% in 2000 (P = 0.005) . Despite an increase in the incidence of nosocomial fungal infection and increased consumption of fluconazole from 1994 to 2000, there was no significant change in the susceptibility to fluconazole for bloodstream isolates of Candida species . These findings appear to be attributed to several factors . These include low prevalence of C . krusei and C . glabrata, changing patterns of use of antifungal drugs and broad-spectrum antibiotics, and efforts to improve the rational use of antifungal agents at our hospital.

FEMS Immunol Med Microbiol, 2003 Jun 10, 37(1), 77 - 83
HIV-1 and its transmembrane protein gp41 bind to different Candida species modulating adhesion; Gruber A et al.; Oral candidiasis in HIV-1-infected individuals is widely believed to be triggered by the acquired T-lymphocyte immunodeficiency . Recently, binding of the HIV-1 envelope protein gp160 and its subunit gp41, and also of the whole virus itself, to Candida albicans has been shown . The present study shows that, in addition to C . albicans, HIV-1 gp41 also binds to yeast and hyphal forms of Candida dubliniensis, a species which is closely related to C . albicans, and to Candida tropicalis but not to Candida krusei, Candida glabrata or Saccharomyces cerevisiae . The previous finding that gp41 binding to C . albicans augments fungal virulence in vitro is supported by the observation that the yeast showed an enhanced adhesion to HIV-infected H9 cells in comparison to uninfected cells . In line with these results soluble gp41 itself reduced binding of C . albicans to both endothelial and epithelial cell lines, confirming a dominant role of the gp41 binding moiety on the surface of Candida for adhesion . Surface-associated secreted aspartic proteinases (Saps) play an important role in candidial adhesion, but are not likely to be involved in the interaction as gp41 binding to the C . albicans parental wild-type strain was comparable to that of three different isogenic Sap deletion mutants . Furthermore, gp41 binding to the yeast killer toxin-susceptible C . albicans strain 10S was not inhibitable by an anti-YKT receptor antibody . In conclusion, HIV-1 interacts with different clinically important Candida spp., and may thereby affect the outcome of the respective fungal infection.

Appl Microbiol Biotechnol, 2003 Jun, 61(5-6), 488 - 94 Epub 2003 Feb 20.
Structure-function study of the amino-terminal stretch of the catalase subunit molecule in oligomerization, heme binding, and activity expression; Ueda M et al.; Analysis of the protein structure of bovine liver catalase suggested that the N-terminal region containing two alpha-helices may function as a linker binding to another subunit . The number of amino-acid residues in catalase from the n-alkane-assimilating yeast Candida tropicalis (CTC) is the lowest of any eukaryotic catalase molecule hitherto investigated, and only one helix, corresponding to the helix alpha2 in bovine liver catalase, is estimated to be present in the same region . In the present study, N-terminal-deleted mutants of CTC were characterized to evaluate the role of the alpha-helix structure in the N-terminal region . CTCDelta1-4 and CTCDelta1-24, whose N-terminal regions were shortened by four and 24 amino-acid residues, respectively, showed an 80% decrease in specific activity compared to wild-type CTC in spite of containing the same amount of heme as in the wild-type . Polyacrylamide gel electrophoresis under nondenaturing conditions revealed that the mutants contained large amounts of oligomeric forms with molecular masses less than 220 kDa (tetramer assembly) . Although the smaller oligomers were found to be bound with heme, only the tetramer exhibited catalase activity in activity staining on nondenaturing gel . CTCDelta1-49, a mutant with deletion of the N-terminal 49 amino-acid residues which contain the conserved helix alpha2, showed no catalase activity and no heme binding . However, the CD spectrum profiles of CTCDelta1-49, CTCDelta1-4, and CTCDelta1-24 indicated that these mutant subunits could attain secondary conformations similar to that of wild-type CTC, regardless of their binding with heme . From these results, it was concluded that the N-terminal stretch of catalase is significant for complete assembly into active tetramer and that the conserved helix alpha2, although it has little effect on the formation of the subunit secondary structure, is indispensable not only in assembling tetramer but also in binding heme.

Harefuah, 2003 Apr, 142(4), 263 - 4, 318
{Presumed Candida tropicalis endophthalmitis following anterior resection}; Schaal S et al.; We present a case of a 75 years old patient who developed a presumed Candida Tropicalis endophthalmitis after an anterior resection for rectal carcinoma . The pathogen was identified in blood cultures but not in vitreal cultures . Systemic anti fungal treatment brought no ocular improvement . After having a vitrectomy and intravitreal antifungal agent injection gradual improvement up to full resolution took place . To the best of our knowledge, this is the first report of a post surgical candida tropicalis endophthalmitis in a non-neutropenic patient . We suggest special attention to ophthalmic complaints in patients with systemic candidemia and prompt treatment according to ocular findings.

Appl Microbiol Biotechnol, 2003 Nov, 63(1), 96 - 100 Epub 2003 May 15.
Effect of redox potential on stationary-phase xylitol fermentations using Candida tropicalis; Kastner JR et al.; Redox potential was used to develop a stationary-phase fermentation of Candida tropicalis that resulted in non-growth conditions with a limited decline in cell viability, a xylitol yield of 0.87 g g(-1) (95% of the theoretical value), and a high maximum specific production rate (0.67 g g(-1) h(-1)) . A redox potential of -100 mV was found to be optimum for xylitol production over the range 0-150 mV {correction} . A shift from ethanol to xylitol production occurred when the redox potential was reduced from 50 mV to 100 mV as cumulative ethanol (Y(ethanol)) decreased from 0.34 g g(-1) to 0.025 g g(-1) and Y(xylitol) increased from 0.15 g g(-1) to 0.87 g g(-1) (alpha=0.05) . Reducing the redox potential to 150 mV did not improve the fermentation . Instead, the xylitol yield and productivity decreased to 0.63 g g(-1) and 0.58 g g(-1) h(-1) respectively and cell viability declined . The viable, stationary-phase fermentation could be used to develop a continuous fermentation process, significantly increasing volumetric productivity and reducing downstream separation costs, potentially by the use of a membrane cell-recycle reactor.

Nippon Ishinkin Gakkai Zasshi, 2003, 44(2), 93 - 100
{Subtractive gene cloning and gene-disruption for elucidation of pseudohyphal formation in Candida tropicalis}; Suzuki T; The dimorphic transition from yeast to pseudohyphae in the petroleum-assimilating yeast Candida tropicalis occurs following the addition of ethanol to glucose semi-defined medium . Subtractive gene cloning was performed on the cDNA from the yeast-growing control culture and on that from the ethanol-supplemented one (the ethanol culture) . A homologue of Schizosaccharomyces pombe nmt1+ or Saccharomyces cerevisiae THI5 was isolated from the cDNA fraction as a preferentially expressed gene for the ethanol culture . This homologue was tentatively called Ctnmt1+, since exogenous thiamine repressed its expression in C . tropicalis growth media . The ethanol culture showed a biphasic pattern of growth phases and the expression of Ctnmt1+ occurred at the first growth phase . The supplementation of thiamine to the ethanol culture at the first phase was followed by repression of Ctnmt1+ expression and also delay of pseudohyphal growth: filamentous growth was inhibited and chains of yeast cells were formed . A Ctnmt1+ disruptant of this organism did not show thiamine auxotrophy and produced pseudohyphal filaments even in the control culture . The supplementation of oxythiamine, an analog of thiamine, to the control culture was followed by the appearance of pseudohyphal filaments, indicating the participation of thiamine during the process of pseudohyphal growth in this organism.

Int J Food Microbiol, 2003 Jun 25, 83(3), 307 - 18
Use of starter cultures of lactic acid bacteria and yeasts in the preparation of togwa, a Tanzanian fermented food; Mugula JK et al.; Starter cultures of lactic acid bacteria (Lactobacillus brevis, Lactobacillus cellobiosus, Lactobacillus fermentum, Lactobacillus plantarum and Pediococcus pentosaceus) and yeasts (Candida pelliculosa, Candida tropicalis, Issatchenkia orientalis and Saccharomyes cerevisiae) isolated from native togwa were tested singly or in combination for their ability to ferment maize-sorghum gruel to produce togwa . All species of bacteria showed an ability to ferment the gruel as judged by lowering the pH from 5.87 to 3.24-3.49 and increasing the titratable acidity from 0.08% to 0.30-0.44% (w/w, lactic acid) in 24 h . Yeasts used singly showed little activity within 12 h, but lowered the pH to 3.57-4.81 and increased the acidity to 0.11-0.21% in 24 h . Yeasts in co-culture with lactic acid bacteria (LAB) had a modest effect on the final acidity (P<0.05) . The number of lactic acid bacteria and yeasts increased while the Enterobacteriaceae decreased with fermentation time . The pH was lowered and lactic acid produced significantly (P<0.05) fastest in natural togwa fermentation and in samples fermented by L . plantarum or L . plantarum in co-culture with I . orientalis . The content of fermentable sugars was reduced during fermentation . Most volatile flavour compounds were produced in samples from fermentation by P . pentosaceus and I . orientalis in co-culture with either L . plantarum or L . brevis.

Kansenshogaku Zasshi, 2003 Mar, 77(3), 158 - 66
{A study for candidemia during the six year period from 1993 to 1999 in St . Luke's International Hospital}; Kazama I et al.; There were 71 patients with candidemia in our hospital from November 1, 1993 to October 31, 1999 . We investigated the 59 patients from isolated species, route of infection, underlying disorders, risk factors, complications, treatment and prognosis . Candida albicans was the most commonly isolated species (52%), followed by Candida tropicalis (11%) . Eighty eight percent of the patients developed candidemia from central venous catheter related infections . The risk factors to candidemia included keeping the catheter in place for more than 5 days, gastrointestinal tract malignancies, postoperative state of gastrointestinal tract surgery, administration of broad-spectrum or combination antibiotics for more than 5 days, and under corticosteroid therapy . About half of the patients (47%) had complications, including endophthalmitis (19 patients, 32%), septic shock (12 patients, 20%) . Mortality rate associated with candidemia was 46% . Mortality rate was lower in 20 patients who were treated with amphotericin B (40%) than in 34 patients treated with only fluconazole (50%), but it was not statistically significant . In order to make an early diagnosis of candidemia, taking blood cultures and ophthalmologic examinations are essential, especially for patients who have those risk factors to candidemia mentioned above . If the patient was suspected of having catheter related infection, the catheter should be removed quickly and the catheter tip should be cultured . Once candidemia is found, ophthalmologic examination and systemic antifungal therapy are needed . Antifungal therapy with Amphotericin B should be used for patients with severe candidemia or with candidemia of non-albicans Candida species.

Ann Hematol, 2003 Apr, 82(4), 231 - 5 Epub 2003 Mar 15.
FLAG-IDA in the treatment of refractory/relapsed acute myeloid leukemia: single-center experience; Pastore D et al.; We evaluated the efficacy and toxicity profiles of the combination of fludarabine, high-dose cytosine arabinoside (AraC), idarubicin, and granulocyte colony-stimulating factor (G-CSF) in refractory/relapsed acute myeloblastic leukemia (AML) patients . Between October 1998 and February 2002, 46 AML patients were treated with FLAG-IDA (fludarabine 30 mg/m(2), AraC 2 g/m(2) for 5 days, idarubicin 10 mg/m(2) for 3 days, and G-CSF 5 micro g/kg from day +6 until neutrophil recovery) . Thirty patients were in relapse after conventional chemotherapy including cytarabine, etoposide, and daunorubicin or mitoxantrone according to the GIMEMA protocols . Four were in relapse after autologous peripheral stem cell transplantation and two after allogeneic bone marrow transplantation . Ten patients had refractory disease (after 10 days of standard doses of cytarabine, 3 days of mitoxantrone or daunorubicin, and 5 days of etoposide) . Recovery of neutrophils and platelets required a median of 19 and 22 days from the start of therapy . Complete remission (CR) was obtained in 24 of 46 patients (52.1%) and 3 of 46 (6.6%) died during reinduction therapy: 2 due to cerebral hemorrhage and 1 due to fungemia ( Candida tropicalis) . Fever >38.5 degrees C was observed in 40 of 46 patients (86.9%), 27 had fever of unknown origin (FUO) and 13 documented infections; 31 of 46 (67.3%) developed mucositis and 14 of 46 (30.4%) had grade 2 WHO transient liver toxicity . After achieving CR, 11 patients received allogeneic stem cell transplantation, 4 patients received autologous stem cell transplantation, 4 were judged unable to receive any further therapy, and 5 refused other therapy . Ten patients are at present in continuous CR after a median follow-up of 13 months (range: 4-24) . In our experience, FLAG-IDA is a well-tolerated and effective regimen in relapsed/refractory AML . The toxicity is acceptable, enabling most patients to receive further treatment, including transplantation procedures.

FEMS Yeast Res, 2003 Mar, 3(1), 3 - 9
Identification of yeast species from orange fruit and juice by RFLP and sequence analysis of the 5.8S rRNA gene and the two internal transcribed spacers; Las Heras-Vazquez FJ et al.; Yeast isolates from orange fruit and juice in a spontaneous fermentation were identified and classified by two molecular techniques . The first was analysis of the restriction pattern generated from the polymerase chain reaction (PCR)-amplified 5.8S rRNA gene and the two internal transcribed spacers (ITS) using specific primers . The second technique was sequence analysis of the ITS regions using the same two primers . Nine different restriction profiles were obtained from the size of the PCR products and the restriction analyses with three endonucleases (CfoI, HaeIII and HinfI) . These groups were identified as Candida tropicalis, Clavispora lusitaniae, Hanseniaspora uvarum, Pichia anomala, Pichia fermentans, Rhodotorula mucilaginosa, Saccharomyces cerevisiae, Saccharomyces unisporus, and Trichosporon asahii . Checking against identification according to morphological, physiological and biochemical traits corroborated this molecular identification . A total concordance was found in the identification with PCR-restriction fragment length polymorphism of the ITS region after analysing certified yeast strains from two different culture collections . Consequently, a rapid and reliable identification of the yeast populations was achieved by using molecular techniques.

Biomedica, 2003 Mar, 23(1), 31 - 7
{Evaluation of Candida species' susceptibility to fluconazole with the disk diffusion method}; de Bedout C et al.; Infections caused by yeasts belonging to the genus Candida have increased dramatically in the last decades, especially in hospital settings . Concomittantly, antimycotic resistance has emerged, as well as the appearance of non-Candida albicans isolates . To standardize in vitro antifungal susceptibility tests, the agar diffusion test was developed using disks impregnated with the antimycotic compound . Electronic recording of the inhibition zone (BIOMIC), furnishes objective values for the minimal inhibitory concentration (MIC) . The fluconazole susceptibility patterns were determined for Candida species isolated from 2.139 patients seen in outpatient clinics or in health-care centers in Colombia, Ecuador and Venezuela . Candida albicans was the species most frequently isolated (62%), followed at a distance by Candida parapsilosis (11%), Candida tropicalis (8.5%), Candida glabata (3.5%) and Candida krusei (2.2%) . MIC determinations showed that 88.1% of these isolates were susceptible to fluconazole, 5.1% were susceptible-dose-dependant and 6.8% resistant . An important proportion (92.1%) of the C . albicans isolates proved susceptible while resistance predominated in the remaining species . These results indicate that the BIOMIC method is rapid and simple, constituting a suitable tool for the epidemiologic surveillance of resistance in Candida species.

Bioresour Technol, 2003 Feb, 86(3), 235 - 7
Washout of a yeast population during continuous treatment of salad-oil-manufacturing wastewater; Zheng S et al.; During continuous treatment of salad-oil-manufacturing wastewater using yeast isolates, a large loss of biomass was observed, which subsequently reduced the treatment efficiency . The correlation between biomass washout and loss of species was monitored using the microbiological characteristics of the effluent and the aeration tank . Of the five yeast species, only Candida tropicalis ultimately remained in the aeration tank possibly because it had the best settleability . Addition of nitrogen to the final effluent stimulated the activity of this yeast, which resulted in a treatment efficiency similar to that of the mixed yeast system.

Sheng Wu Gong Cheng Xue Bao, 2002 Nov, 18(6), 724 - 8
{Utilization of sugar cane bagasse hydrolysates for xylitol production by yeast}; Zhang HR et al.; The effects of the concentration of sulfuric acid and the ratio of liquid to solid on xylose yield from sugar cane bagasse in its hemicellulose hydrolysis process were studied with the Quadratic Rotary Combination Design . Regression analysis showed that there was a marked regression relationship between the two factors and xylose yield . As the result of optimizing the hydrolysis conditions by regression equation, xylose yield of 24 g/100 g sugar cane bagasse was obtained when sulfuric acid concentration was 2.4 g/L and liquid to solid ratio was 6.2 under the conditions of stream pressure of 2.5 x 10(4) Pa and hydrolysis time of 2.5 h . The macroporous resin adsorption was proved to be a good method to reduce the concentration of yeast cell growth inhibitor in sugar cane bagasse hemicellulose hydrolysate and to enhance the hydrolysate fermentability . The hydrolysate treated with macroporous resin adsorption under pH2 was used as the substrate for xylitol production by a xylitol-producting yeast, Candida tropicalis AS2.1776 . At an initial xylose concentration of 200 g/L, all xylose was consumed within 110 h with a xylitol production rate of 1.15 g/L.h, and a xylitol yield of 0.64 g/g xylose.

Indian Pediatr, 2003 Mar, 40(3), 261 - 4
Drug resistant neonatal Candida tropicalis septicemia . Did it cause diaphragmatic hernia?
Dutta S, Narang A.
A full-term, 3 kg baby girl developed early onset Candida tropicalis septicemia . The fungus was resistant to amphotericin B, fluconazole and itraconazole . She developed an acquired diaphragmatic hernia during the course of the infection . The possible association of the hernia with the fungal sepsis is discussed . She improved on treatment with 4-flucytosine and after being operated for the hernia.

J Infect, 2003 Apr, 46(3), 155 - 60
Candida tropicalis fungemia in a tertiary care hospital; Goldani LZ et al.; Candida tropicalis is a frequent cause of fungemia in hospitals in Latin America . Candida albicans (33%) was the most frequently isolated species, followed by Candida parapsilosis (27%), and Candida tropicalis (24%) in tertiary care hospital in Brazil . We identified and retrospectively reviewed 27 cases of C . tropicalis fungemia that occurred at Hospital de Clinicas de Porto Alegre from 1996 to 1999 . The mean age of the patients was 32 years (range 6 months to 88 years) . Eight patients (29.6%) had hematological malignancy, and four (14.8%) had solid tumors . All the patients were taking broad-spectrum antibiotics, including vancomycin for at least 7 days . Antibiotics were given through a central venous catheter for the majority of the patients (77.7%) . Relevant risk factors for candidemia in our patients included neutropenia (59.2%), and use of corticosteroids (37.0%) or cytotoxic drugs (40.7%) . The onset of fever was the most frequent clinical manifestation (92.5%) of fungemia . Most of the patients (81.4%) were treated with amphotericin B or fluconazole . Overall mortality was 48.1%, and 7 (53.4%) of 13 deaths occurred within 10 days of the detection of candidemia . Results of the in vitro susceptibility testing of nine isolates of C . tropicalis from seven patients did not show resistance to fluconazole and amphotericin B.C . tropicalis presents as an important cause of fungemia in oncological and nononcological patients with central venous catheters taking broad-spectrum antibiotics . Although there was no evidence of resistance of C . tropicalis to amphotericin B and fluconazole, patients treated with antifungal agents presented with a high mortality rate in the hospital setting.

Biochim Biophys Acta, 2003 Mar 17, 1631(2), 160 - 8
Up-regulation of the peroxisomal beta-oxidation system occurs in butyrate-grown Candida tropicalis following disruption of the gene encoding peroxisomal 3-ketoacyl-CoA thiolase; Ueda M et al.; In the yeast Candida tropicalis, two thiolase isozymes, peroxisomal acetoacetyl-CoA thiolase and peroxisomal 3-ketoacyl-CoA thiolase, participate in the peroxisomal fatty acid beta-oxidation system . Their individual contributions have been demonstrated in cells grown on butyrate, with C . tropicalis able to grow in the absence of either one . In the present study, a lack of peroxisomal 3-ketoacyl-CoA thiolase protein resulted in increased expression (up-regulation) of acetoacetyl-CoA thiolase and other peroxisomal proteins, whereas a lack of peroxisomal acetoacetyl-CoA thiolase produced no corresponding effect . Overexpression of the acetoacetyl-CoA thiolase gene did not suppress the up-regulation or the growth retardation on butyrate in cells without peroxisomal 3-ketoacyl-CoA thiolase, even though large amounts of the overexpressed acetoacetyl-CoA thiolase were detected in most of the peroxisomes of butyrate-grown cells . These results provide important evidence of the greater contribution of 3-ketoacyl-CoA thiolase to the peroxisomal beta-oxidation system than acetoacetyl-CoA thiolase in C . tropicalis and a novel insight into the regulation of the peroxisomal beta-oxidation system.

Folia Microbiol (Praha), 2002, 47(6), 701 - 7
Kinetics of phenol oxidation by Candida tropicalis: effects of oxygen supply rate and nutrients on phenol inhibition; Paca J et al.; The kinetics of phenol degradation was estimated in a fed-batch reactor system . Effects of oxygen and nutrient excess or limitation as well as the presence of several essential ions on the phenol- and oxygen-specific uptake rates achieved simultaneously in a bioreactor were shown . Candida tropicalis was grown on phenol as the only carbon and energy source . Applying the best fit of polynomial function, the maximum specific uptake rates of phenol and oxygen, the critical concentrations of phenol, the half-saturation constants and inhibition constants were determined . Linear relationship between specific phenol uptake rate and the exogenous respiration rate was found regardless of the kind and presence of essential nutrients . At oxygen limitation both the phenol uptake rate and the cell affinity to phenol decreased more strongly compared with those under nutrient limitation . Oxygen in excess resulted in a significant increase of cell tolerance toward phenol . The presence of essential nutrients increased the specific phenol degradation rate and led to complete phenol oxidation.

J Med Microbiol, 2003 Mar, 52(Pt 3), 229 - 38
Detection of seven Candida species using the Light-Cycler system; White PL et al.; Due to the limitations of classical methods for the detection of systemic fungal infections and the high mortality rates associated with these infections, it has become essential to develop a quick, sensitive and specific detection assay . By using the Idaho Technologies Light-Cycler system, a qualitative real-time PCR system has been developed for the detection of the leading causes of systemic infection within the genus CANDIDA: The sensitivity of the assay was comparable to previously described PCR methods (1-5 c.f.u . ml(-1)) and, by the use of a single Candida probe, it was able to detect, but not differentiate between, seven species of Candida (Candida albicans, Candida dubliniensis, Candida glabrata, Candida kefyr, Candida krusei, Candida parapsilosis and Candida tropicalis) . Single-round amplification on the Light-Cycler allowed rapid turn-around of clinical samples (within one working day) and it was shown to be more sensitive than classical procedures, exposing 39 possible systemic infections that were not detected by blood culture.

Metab Eng, 2002 Jul, 4(3), 248 - 56
Metabolic flux analysis of Candida tropicalis growing on xylose in an oxygen-limited chemostat; Granstrom T et al.; We have studied the metabolism of xylose by Candida tropicalis in oxygen-limited chemostat . In vitro enzyme assays indicated that glycolytic and gluconeogenetic enzymes are expressed simultaneously facilitating substrate cycling . Enhancing the redox imbalance by cofeeding of formate increased xylose and oxygen consumption rates and ethanol, xylitol, glycerol and CO2 production rates at steady state . Metabolic flux analysis (MFA) indicated that fructose 6-phosphate is replenished from the pentose phosphate pathway in sufficient amounts without contribution of the gluconeogenetic pathway . Substrate cycling between pyruvate kinase, pyruvate carboxylase and phospho-enol-pyruvate kinase increased ATP turnover . Cofeeding of formate increased the ATP yield . The ATP yields of xylose and xylose-formate cultivation were 6.9 and 8.7 mol ATP/C-mol CDW, respectively, as calculated from the MFA.

J Mol Biol, 2003 Mar 14, 327(1), 47 - 59
Structure-function analysis of enoyl thioester reductase involved in mitochondrial maintenance; Airenne TT et al.; Candida tropicalis enoyl thioester reductase Etr1p and the Saccharomyces cerevisiae homologue Mrf1p catalyse the NADPH-dependent reduction of trans-2-enoyl thioesters in mitochondrial fatty acid synthesis (FAS) . Unlike prokaryotic enoyl thioester reductases (ETRs), which belong to the short-chain dehydrogenases/reductases (SDR), Etr1p and Mrf1p represent structurally distinguishable ETRs that belong to the medium-chain dehydrogenases/reductases (MDR) superfamily, indicating independent origin of two separate classes of ETRs . The crystal structures of Etr1p, the Etr1p-NADPH complex and the Etr1Y79Np mutant were refined to 1.70A, 2.25A and 2.60A resolution, respectively . The native fold of Etr1p was maintained in Etr1Y79Np, but the mutant had only 0.1% of Etr1p catalytic activity remaining and failed to rescue the respiratory deficient phenotype of the mrf1Delta strain . Mutagenesis of Tyr73 in Mrf1p, corresponding to Tyr79 in Etr1p, produced similar results . Our data indicate that the mitochondrial reductase activity is indispensable for respiratory function in yeast, emphasizing the significance of Mrf1p (Etr1p) and mitochondrial FAS for the integrity of the respiratory competent organelle.

Ann Hematol, 2003 Feb, 82(2), 93 - 7 Epub 2003 Jan 08.
Hepatosplenic fungal infection in patients with acute leukemia in Taiwan: incidence, treatment, and prognosis; Chen CY et al.; Nosocomial fungal infection increases gradually and has become the leading pathogen at National Taiwan University Hospital since 1993 . From January 1995 through May 2002, hepatosplenic fungal infection (HSF) was diagnosed in 37 (7.4%) of the 500 adult patients with acute leukemia who received chemotherapy at this hospital . There was no significant difference in the incidence of HSF between the patients with acute myeloid leukemia and those with acute lymphoblastic leukemia, or between the patients treated with high-dose chemotherapy and those with conventional or low-dose chemotherapy . Candida tropicalis was the leading pathogen, followed by Candida albicans . The computed tomography scan showed multiple hypodense lesions in the liver (89%), spleen (70%), and kidney (27%) . Eighteen patients were initially treated with fluconazole and 19 with amphotericin B . Nineteen patients received the planned chemotherapy after the diagnosis of HSF . Among them, eight patients underwent hematopoietic stem cell transplantation and seven patients survived more than 100 days post-transplantation; none of these patients had relapse of prior HSF . Twenty-three patients (62%) died during a median follow up of 10 months, but only seven died due to HSF . In conclusion, a substantial percentage of patients with acute leukemia acquired HSF after chemotherapy and carried high mortality . However, HSF itself is not a contraindication for subsequent chemotherapy and hematopoietic stem cell transplantation.

FEMS Microbiol Lett, 2003 Feb 14, 219(1), 93 - 8
Effect of catalase-specific inhibitor 3-amino-1,2,4-triazole on yeast peroxisomal catalase in vivo; Ueda M et al.; 3-Amino-1,2,4-triazole (3-AT) is known as an inhibitor of catalase to whose active center it specifically and covalently binds . Subcellular fractionation and immunoelectronmicroscopic observation of the yeast Candida tropicalis revealed that, in 3-AT-treated cells in which the 3-AT was added to the n-alkane medium from the beginning of cultivation, catalase transported into peroxisomes was inactivated and was present as insoluble aggregated forms in the organelle . The aggregation of catalase in peroxisomes occurred only in these 3-AT-treated cells and not in cells in which 3-AT was added at the late exponential growth phase . Furthermore, 3-AT did not affect the transportation of catalase into peroxisomes . The appearance of aggregation only in cells to which 3-AT was added from the beginning of cultivation suggests that, in the process of catalase transportation into yeast peroxisomes, some conformational change may take place and that correct folding may be inhibited by the binding of 3-AT to the active center of catalase . Accordingly, 3-AT will be an interesting compound for investigation of the transport machinery of the peroxisomal tetrameric catalase.

J Clin Microbiol, 2003 Feb, 41(2), 735 - 41
Candida tropicalis in a neonatal intensive care unit: epidemiologic and molecular analysis of an outbreak of infection with an uncommon neonatal pathogen; Roilides E et al.; From June to July 1998, two episodes of Candida tropicalis fungemia occurred in the Aristotle University neonatal intensive care unit (ICU) . To investigate this uncommon event, a prospective study of fungal colonization and infection was conducted . From December 1998 to December 1999, surveillance cultures of the oral cavities and perinea of the 593 of the 781 neonates admitted to the neonatal ICU who were expected to stay for >7 days were performed . Potential environmental reservoirs and possible risk factors for acquisition of C . tropicalis were searched for . Molecular epidemiologic studies by two methods of restriction fragment length polymorphism analysis and two methods of random amplified polymorphic DNA analysis were performed . Seventy-two neonates were colonized by yeasts (12.1%), of which 30 were colonized by Candida albicans, 17 were colonized by C . tropicalis, and 5 were colonized by Candida parapsilosis . From December 1998 to December 1999, 10 cases of fungemia occurred; 6 were due to C . parapsilosis, 2 were due to C . tropicalis, 1 was due to Candida glabrata, and 1 was due to Trichosporon asahii (12.8/1,000 admissions) . Fungemia occurred more frequently in colonized than in noncolonized neonates (P < 0.0001) . Genetic analysis of 11 colonization isolates and the two late blood isolates of C . tropicalis demonstrated two genotypes . One blood isolate and nine colonization isolates belonged to a single type . The fungemia/colonization ratio of C . parapsilosis (3/5) was greater than that of C . tropicalis (2/17, P = 0.05), other non-C . albicans Candida spp . (1/11, P = 0.02), or C . albicans (0/27, P = 0.05) . Extensive environmental cultures revealed no common source of C . tropicalis or C . parapsilosis . There was neither prophylactic use of azoles nor other risk factors found for acquisition of C . tropicalis except for total parenteral nutrition . A substantial risk of colonization by non-C . albicans Candida spp . in the neonatal ICU may lead to a preponderance of C . tropicalis as a significant cause of neonatal fungemia.

J Antimicrob Chemother, 2003 Feb, 51(2), 297 - 304
Comparison of three methods for in vitro susceptibility testing of Candida species with flucytosine; Moore CB et al.; Optimal methods for susceptibility testing of Candida spp . with flucytosine have not been determined . Breakpoints were recommended in 1984, but never validated . In this study, we compared the 1984 recommended macrodilution broth method (using an 80% endpoint) with a modification of the more recent NCCLS-recommended microdilution broth method with three endpoints-spectrophotometric 50% and 80% and a no growth endpoint determined by eye . NCCLS and British Society for Medical Mycology (BSMM) breakpoints were also compared . One hundred and fifty isolates comprised of Candida albicans, Candida tropicalis, Candida krusei, Candida glabrata, Candida parapsilosis and Candida lusitaniae were tested . Reproducibility was excellent . For C . albicans (n = 65), the correlation between tests was excellent (>75%), with few major discrepancies (<5%) . For C . tropicalis (n = 27), correlation was good (59%), but there were a small number of major discrepancies (up to 11%, depending on breakpoint used) . Results by the broth macrodilution method were generally higher than both microdilution methods for C . glabrata (n = 16; correlation of 18.8%), but only one major discrepancy was seen . Ten of the 11 C . parapsilosis isolates tested were susceptible by all methods, regardless of breakpoint chosen, with a correlation of 18.2%, but no major discrepancies were seen . A correlation between all methods (50%) was seen with C . lusitaniae (n = 10), with many isolates resistant or intermediate . In contrast, correlation between methods for C . krusei was poor (<5%); NCCLS microtitre modification produced results that were classified as intermediate or resistant, regardless of the breakpoint used . The methodology for susceptibility testing C . albicans is robust . Additional work to optimize susceptibility testing with flucytosine is necessary for non-albicans Candida species, especially C . krusei.

Wei Sheng Wu Xue Bao, 2002 Apr, 42(2), 193 - 9
{Analysis of POX4 and POX5 gene encoded proteins of Candida tropicalis 1230}; Qin W et al.; POX4 and POX5 were cloned from Candida tropicalis with a simple method . The sequencing results show differences of POX5 between strain 120 and strain pk233 . The searching of these two sequences against the curreent collection of Pfam profile HMMS located regions that belong to known domain families . Secondary structure prediction shows that N-terminal secondary structure of PXP4 and PXP5 are homologous to that of pig liver medium-chain acyl-coA dehydrogenase . Thus, the N-terminal of PXP4 and PXP5 may be involved in FAD binding.

Wei Sheng Wu Xue Bao, 2002 Jun, 42(3), 359 - 63
{Effects of H2O2 addition on cell growth and product formation in long-chain dicarboxylic acid fermentation}; Li S et al.; When Candida tropicalis was cultivated in shakig-flask with the H2O2 addition, DCA (Dicarboxylic Acid, DCA) concentration was increased, especially at 2 mmol/L H2O2 concentration . The cytochrome P450 activity assays indicated that H2O2 addition significantly increased the activities of cytochrome P450 and DCA production positively correlated with the activities of cytochrome P450 . The study on the cell growth demonstrated that the H2O2 addition inhibited the cell growth rate . However, the retarding effect was not irreversible since the cell growth rate could recover slowly to the original level after the H2O2 addition was halted . The mechanism of inducement on cytochrome P450 by H2O2 addition was also discussed in this article.

Wei Sheng Wu Xue Bao, 2002 Feb, 42(1), 114 - 6
{Study on fermentation of n-paraffin for producing mixed dicarboxylic acids}; Tong M et al.; A mutant of Candida tropicalis FYD-2 was obtained from its parental strain SFP-1186 by ultraviolet treatments . On shaking flask, the yield of mixed dicarboxylic acid(DCA) by the mutant was 21.4% higher than that by its ancestor . The amount of mixed DCA reached 156 g/L for 120 h incubation in a 10 L autoconrolled fermentor where the culture medium contained 25% n-paraffin . The process of induced and screening mutant was introduced and the time course of fermentation in 10 L fermentor was discussed.

Wei Sheng Wu Xue Bao, 1999 Jun, 39(3), 279 - 81
{Studies on microbial production of undecane 1, 11-dicarboxylic acid from N-tridecane}; Chen Y et al.; A mutant, Candida tropicalis P-12-242, which can produce undecane 1, 11-Dicarboxylic acid(DC13) from N-Tridecane(nC13), was obtained by treating the parent stain UH-2-48 with sodium nitrite . On 2500L fermenter testing, under the optimum condition where the fermentation medium contained total 26% nC13, pH of the course of fermentation was maintained range 7.5-8.0, at 28 degrees C-30 degrees C, the highest level of DC13 production was obtained after 6 d, and the average amount of DC13 accumulated was 182.6 g/L in broth . After received residual nC13, The average consumption rate of DC13 from nC13 was 87.8% . The purity of the product DC13, which was analyzed by gas chromatography was about 96.8%.

Wei Sheng Wu Xue Bao, 1998 Aug, 38(4), 318 - 20
{Screening and breeding of highly-effected degrading cotton-phenol strains and study on defoxication technology and conditions}; Shi A et al.; From mildewed cottonseed cake and stock cultures of mold and yeast . We select more than ten strains of yeasts and molds which can degrade cotton phenol . At last we got four strains which can degrade cotton phenol highly effected after mutagenized by physical and chemical factors and induced by cotton phenol . They belong to Candida tropicalis, Torulopsis candida, Aspergillus flavus and Aspergillus niger . By small and medium size fermenfations, the content of dissociated cotton phenol all reach safe criterion (the lowest dissociated cotton phenol content is 220 mg/L in the defoxicated cottonseed cake), compared with FeSO4 method, defoxicated cottonseed cake looks and tastes well and the content of protein and amino acid are highly enhanced as will.

Wei Sheng Wu Xue Bao, 2001 Feb, 41(1), 117 - 20
{Study on the cytochrome P450 activity in alkane converting process of Candida tropicalis}; Jiao P et al.; A method of reduced CO-difference spectrum was established to study the cytochrome P450 activity of the whole cell of Candida tropicalis during the alkane converting process . Using this method, the cytochrome P450 activities of the whole cells that were cultured in the different concentrations of alkane were studied . The results showed that the 5% alkane could induce the cytochrome P450 activity obviously but not inhibit the growth of cells, so it was determined preliminarily that the alkane concentration of the seed medium was 5% . The cytochrome P450 activities of dicarboxylic acid (DCA) fermentation processing were further studied . During the exponential phase of growth, the cytochrome P450 activity increased smoothly . However, during the phase of production of dicarboxylic acid, the cytochrome P450 activities increased rapidly after a sort decrease . The results still showed that the rate of production of dicarboxylic acid increased with the cytochrome P450 activity.

Wei Sheng Wu Xue Bao, 2000 Jun, 40(3), 318 - 22
{Study on fermentation of 1,13-tridecanedioic acid by Candida tropicalis}; Liu S et al.; A mutant of Candida tropicalis SP-1, SP-UV-56, which can produce 1,13-tridecanedioic acid or 1,11-dicarboxylic acid (DCA13) 1.25 times as much as its parental strain SP-1, but hardly assimilates nalkane as carbon source for its growth, was obtained by ultraviolet treatments . A fermentation technology with a supplement of acetate at the latter middle of logarithmic growth was developed . Using sucrose as carbon source for its growth and supplying acetate the yield of DCA13 reached 153 g/L for 144 h incubation in a 13.7 L auto-controlled stirred tank, about 29.7% higher than that obtained in the broth without acetate . An effective technique that can maintain higher yield was provided by enhancing mixing of broth and controlling low DO level . With these techniques the DCA13 yield reached 172 g/L in a 20 m3 stirred tank, and produced 2.25 tons DCA13 in 15.0 m3 broth.

Wei Sheng Wu Xue Bao, 2000 Apr, 40(2), 214 - 6
{Effect of sugars and pH on the fermentation of decane 1,10-dicarboxylic acid}; Ren G et al.; Some sugars, such as lactose, maltose, raffinose, glucose etc . can increase the purity of DC12 compared with sucrose . The value of pH can affect the process of producing DC12 . Controlling the pH value between 7.5-8.0 can highly improve the production of DC12 . Candida tropicalis can symport the DC12 with H+ to the outside of the cell.

Trans Am Ophthalmol Soc, 2002, 100, 67 - 70; discussion 70-1
Endophthalmitis in patients with disseminated fungal disease; Feman SS et al.; BACKGROUND/PURPOSE: Fungal endophthalmitis caused by dissemination from extraocular fungal infections has been reported to vary between 9% and 45% . However, recent clinical experience disagrees with that . This study is an investigation of patients in an inner city teaching hospital, the risks associated with endogenous fungal endophthalmitis, and this incidence . METHODS: All ophthalmology consultations between February 1995 and August 2000 that might be associated with disseminated fungal infection were examined in a prospective manner . Patients were excluded if there was no evidence of a positive fungal culture from any site at any time . Visual symptoms were recorded along with ophthalmologic and systemic examination features . Information was gathered, including the identity of cultured organisms, the sites from which the organisms were obtained, and the patients' disposition . RESULTS: During this interval, 170 consultation requests contained the words "endophthalmitis" or "retinitis" and/or indicated concern about disseminated fungal infections . Extraocular fungal infections were found in 114 patients, but only 82 of them had evidence of systemic dissemination . Some patients had more than one organism . The following are listed in decreasing frequency of occurrence: Candida albicans, Torulopsis glabrata, Candida tropicalis, Candida parapsilosis, Candida krusei, Aspergillus niger, and others . Only two patients had evidence of chorioretinitis and progressed to fungal endophthalmitis . CONCLUSIONS: Endophthalmitis was rare among these patients with known fungal infections . Less than 2% had any related ophthalmic manifestations . Nevertheless, since treatment can save vision, evidence of intraocular infection should be sought as eagerly as before.

J Med Microbiol, 2003 Feb, 52(Pt 2), 169 - 71
Antifungal properties of 5-hydroxytryptamine (serotonin) against Candida species in vitro; Lass-Florl C et al.; In this study the minimal inhibitory concentration (MIC) and minimal fungicidal concentration (MFC) of 5-hydroxytryptamine (5 HT, serotonin) against clinical isolates of Candida albicans (n = 11), Candida glabrata (n = 9), Candida tropicalis (n = 10) and Candida parapsilosis (ATCC 22019) using a broth microdilution test were investigated . In addition, it was examined whether delayed regrowth as a post-antifungal effect results following short exposure to 5 HT . 5 HT showed antifungal activity towards all isolates of Candida spp . The isolates yielded comparable MIC and MFC values of 5 HT in the range 0.91-7.34 mM and 1.83-14.68 mM, respectively . A lag in regrowth was dependent on the concentration tested . Treatment for 3 h at concentrations of 5 HT below and equipotent to the MFC resulted in a delayed regrowth of 8-12 h for isolates of Candida spp . In conclusion, these in vitro studies clearly demonstrate antifungal effects of 5 HT . Identifying the mode of action could be of great help in developing and researching new antifungal drugs.

J Gen Appl Microbiol, 1997 Oct, 43(5), 265 - 272
Ascomycetous yeasts from tropical intertidal dark mud of southeast Brazilian estuaries; Soares CA et al.; Four different intertidal estuarine sediments had distinct yeast communities . One-hundred-ninety-three yeast isolates were classified in 47 species, with 34 of these in the genus Candida . Candida tropicalis was the only ascomycetous species isolated from all four sites . Other opportunistic pathogens including Candida glabrata, Candida guilliermondii, Candida parapsilosis and Candida krusei were present, especially at the more polluted sites . Pichia species were also frequent isolates with Pichia membranaefaciens, and its anamorph, Candida valida, and other phenotypically similar low assimilation profile species the most frequent . Kluyveromyces aestuarii was prevalent at the only site with well established mangrove vegetation, but not present at the other sites . The sediment yeast communities were distinct from each other, but more similar to each other than to the yeast communities of other ecosystems in the same geographic region.

J Antimicrob Chemother, 2003 Jan, 51(1), 163 - 6
Antifungal activity of the echinocandin anidulafungin (VER002, LY-303366) against yeast pathogens: a comparative study with M27-A microdilution method; Arevalo MP et al.; This study further evaluated the in vitro activity of anidulafungin (VER002, Versicor Inc.) (LY303366) against 460 clinical yeast isolates . MICs of anidulafungin, fluconazole and itraconazole were determined by following the NCCLS M27-A guidelines . Minimum fungicidal concentrations (MFCs) of anidulafungin were determined for 230 isolates of Candida spp . The activity of anidulafungin in vitro was significantly superior (P < 0.05) to those of itraconazole and fluconazole against Candida albicans, Candida tropicalis, Candida glabrata and Candida krusei, but anidulafungin was less active for Candida famata and Candida parapsilosis . The differences were not significant for the other species evaluated.

J Gen Appl Microbiol, 2000 Oct, 46(5), 245 - 249
Isolation and enzyme determination of Candida tropicalis mutants for DCA production; Jiao P et al.; Techniques, named two-step enrichment and double-time replica-plating method (TEDR), are described that allow a mutated population of Candida tropicalis to be enriched efficiently for mutants deficient in the alkane degradation pathway (Alk(-)) and to be selected easily for mutants increasing in the DCA (dicarboxylic acids) excretion pathway . After C . tropicalis was mutated with ethyl methane sulphonate and ultraviolet, the Alk(-) mutants were enriched (the first step enrichment, up to eightfold in one round of enrichment) by treatment with nystatin in medium SEL1-1 . The mutagen-treated cells were then cultured in medium YPD containing chlorpromazine for further enriching (the second-step enrichment, up to threefold in one round) the mutants with an increasing capacity of alpha- and omega-oxidation . On the other hand, the Alk(-) mutants were readily isolated by the SEL1 replica-plating method by using alkane or glucose as the sole carbon source . A total of 43 Alk(-) mutants were isolated from 2x10(8) mutagen-treated cells . In the following steps, by using SEL2 replica plating, the screening studies showed that of the 43 Alk(-) mutants, 11 strains could accumulate DCA greatly from alkane, and strains 1-12 and 1-3, especially, could produce nearly three times as much DCA as the wild-type organism could . The results showed that the strains had more cytochrome P450 activity and a higher converting capacity of alkane.

Chemotherapy, 2002 Dec, 48(5), 224 - 31
Comparison of in vitro antifungal activities of amphotericin B lipid complex with itraconazole against 708 clinical yeast isolates and opportunistic moulds determined by National Committee for Clinical Laboratory Standards methods M27-A and M38-P; Carrillo-Munoz AJ et al.; We compared the in vitro antifungal activity of amphotericin B lipid complex (ABLC) with that of itraconazole (ITZ) against 535 yeast strains and 173 opportunistic filamentous fungi by using a microdilution method (National Committee for Clinical Laboratory Standards M27-A and M38-P) . The overall geometric mean MIC was 0.13 microg/ml and 0.177 microg/ml for ITZ and ABLC, respectively, and the MIC(50) was 0.125 microg/ml for both agents against yeast isolates . ITZ had a similar or slightly superior efficacy compared to ABLC when tested against Candida albicans, Candida parapsilosis, Cryptococcus neoformans, Candida krusei, Candida glabrata and Candida tropicalis . Effectiveness against C . glabrata was lower for ITZ (MIC(90) 2 microg/ml, and for ABLC, 0.5 microg/ml) . For Aspergillus fumigatus, activity of ITZ was superior in comparison with ABLC (MIC(90) 1 and 16 microg/ml, respectively); MIC(90) for Aspergillus niger was 4 and 2 microg/ml for ABLC and ITZ, respectively . Scedosporium spp . showed a low susceptibility to both ABLC and ITZ . In conclusion, ABLC and ITZ are useful alternatives for the treatment of severe fungal infections . The selection of an antifungal agent should be made considering the toxicological and pharmacological properties and cost/benefit relationship and be supported by the susceptibility of the isolate .

Appl Microbiol Biotechnol, 2002 Dec, 60(4), 469 - 74 Epub 2002 Oct 24.
Long anchor using Flo1 protein enhances reactivity of cell surface-displayed glucoamylase to polymer substrates; Sato N et al.; We investigated the influence of anchor length on the reactivity to polymer substrate of enzyme displayed on yeast cell surfaces . Using various lengths {42, 102, 146, 318, 428, and 1,326 amino acids (aa)} of the C-terminal region of the Saccharomyces cerevisiae Flo1 protein (Flo1p), which plays a major role in yeast flocculation, six display systems with various anchor lengths were constructed . In these systems, the target protein was displayed on the yeast cell surface under the control of the 5'-upstream region of the isocitrate lyase gene of Candida tropicalis ( UPR-ICL) . Cell-surface display of Rhizopus oryzae glucoamylase by these systems was induced and confirmed in all systems by immunofluorescence microscopy and immunoblotting . Flow-cytometer measurement of the fluorescence intensity of immunofluorescence-labeled yeast cells displaying glucoamylase indicated that glucoamylase displayed with longer anchors, especially those of 428 and 1,326 aa in length, had higher reactivity to antibodies . The reactivity of starch to displayed glucoamylase, which was evaluated by plate assay, increased with anchor length, as did the cell growth-rate in starch-containing medium . These results indicate that cell-surface display systems using 428- and 1,326-aa length anchors of Flo1p are effective for the display of enzymes on the outer surface of yeast cells.

Eur J Clin Microbiol Infect Dis, 2002 Nov, 21(11), 767 - 74 Epub 2002 Oct 31.
Candidemia at a tertiary-care hospital: epidemiology, treatment, clinical outcome and risk factors for death; Viudes A et al.; The demographic, clinical and microbiological data of patients with candidemia at the "Hopital Universitario La Fe", a tertiary-care hospital in Valencia, Spain, from 1995 to 1997 was analyzed retrospectively . Candida spp . were isolated in blood cultures from 145 patients, 32% of whom were children (25% of these were neonates) . The most common species isolated was Candida albicans, followed by Candida parapsilosis, Candida krusei and Candida tropicalis . Risk factors for candidemia included underlying disease, therapy with broad-spectrum antibiotics and the presence of a central venous catheter . The majority of children were treated with amphotericin B, whereas 52% of adults received fluconazole . Overall mortality was 44% (30% in children and 50% in adults), and attributable mortality was 30% (24% in children and 33% in adults) . Multivariate analysis indicated that neutropenia, corticosteroid therapy, lack of antifungal treatment, and failure to replace the central venous catheter were factors associated with candidemia-related death . Among the adult population, an APACHE II score greater than 15 predicted candidemia-related death.

J Antimicrob Chemother, 2002 Dec, 50(6), 1071 - 4
In vivo activity of micafungin in a persistently neutropenic murine model of disseminated infection caused by Candida tropicalis; Warn PA et al.; Micafungin is a new echinocandin with broad-spectrum in vitro and in vivo antifungal activity against both Aspergillus and Candida species . We compared the activity of micafungin with that of amphotericin B and fluconazole in a persistently immunocompromised murine model of disseminated candidiasis against a strain of Candida tropicalis that was resistant to amphotericin B and fluconazole in vitro . Mice were rendered persistently neutropenic with multiple doses of cyclophosphamide and infected intravenously with C . tropicalis . Mice were treated with either intraperitoneal amphotericin B (0.5-5 mg/kg per dose), oral fluconazole (50 mg/kg twice a day), intravenous micafungin (1-10 mg/kg per dose) or solvent control for 7 days . Mice were killed at 11 days post-infection and kidneys, lungs, brain and liver removed for quantitative culture . Overall mortality in the model was low, with rates varying between 10% and 25% in treatment groups . Micafungin at doses between 2 and 10 mg/kg were the only regimes able to reduce cfu below the level of detection of tissues infected with C . tropicalis . Micafungin was well tolerated by the mice and was much more effective than amphotericin B or fluconazole against an amphotericin B- and fluconazole-resistant C . tropicalis.

J Clin Microbiol, 2002 Dec, 40(12), 4768 - 70
Persistence of pigment production by yeast isolates grown on CHROMagar Candida medium; Hospenthal DR et al.; We evaluated the persistence of pigmentation in yeast isolates grown on the chromogenic medium CHROMagar Candida over 7 days . Candida, Cryptococcus, and Trichosporon isolates were inoculated alone or mixed onto duplicate sets of plates and incubated at 30 and 35 degrees C . Candida albicans and Candida krusei were readily identified throughout the reading period, but Candida glabrata was difficult to differentiate from other species until the 3- or 4-day time point . Candida tropicalis produced colonies similar to those of rare Cryptococcus and Trichosporon species, and mixed cultures were often difficult to identify as such.

New Microbiol, 2002 Oct, 25(4), 489 - 94
Identification and antifungal susceptibility of Candida isolated from intensive care unit patients; Zer Y et al.; Candida was isolated in 205 of 1060 clinical specimens (19.33%) in our laboratary sent from the intensive care unit for mycological investigation between January 98-December 99 . All isolated strains were identified to species level using the API Candida system (Bio-Meieux, France) as follows; Candida albicans (n:115, 56.09%), Candida tropicalis (n:23, 11.21%), Candida parapsilosis (n:21, 10.24%), Candida glabrata (n:12, 5.83%) . Candida kefyr (n:9, 4.39%), Candida lusitaniae (n:7, 3.41%), Candida famata (n:6, 2.92%), Candida krusei (n:6, 2.92%), Candida guilliermondii (n:6, 2.92%) . These stains were identified using congo-red-glucose-brain-heart-infusion agar and slime production was determined in Candida albicans 53.91% and 67.77% in other than Candida species . In the present study, E test (AB Biodisk, Solna, Sweeden) was used to test antifungal susceptibility . The resistance to amphotericin B was 19.51%, to fluconazole 27.31% and to flucytosine 20.00%.

Antimicrob Agents Chemother, 2002 Dec, 46(12), 3846 - 53
In vitro activity of micafungin (FK-463) against Candida spp.: microdilution, time-kill, and postantifungal-effect studies; Ernst EJ et al.; We evaluated the in vitro activity of the new echinocandin antifungal micafungin against Candida spp . using microdilution and time-kill methods . Additionally, we examined the postantifungal effect (PAFE) of micafungin . Finally, we evaluated the effect of the addition of serum and plasma on the MIC of micafungin . Four Candida albicans isolates and two isolates of each Candida glabrata, Candida krusei, and Candida tropicalis were selected for testing . The MICs of micafungin were determined in RPMI 1640 medium buffered with morpholinepropanesulfonic acid alone and with the addition of 10, 20, and 50% human serum and plasma . MICs were determined by using two endpoints: a prominent reduction in growth (the MIC at which 80% of isolates are inhibited {MIC(80)}) and complete visual inhibition of growth (MIC(100)) . The minimum fungicidal concentration (MFC) of micafungin for each isolate was also determined . Time-kill curves were determined for each isolate in RPMI 1640 medium with micafungin at concentrations ranging from 0.125 to 16 times the MIC(80) to assess the correlation between MIC(80) and fungicidal activity . PAFE studies were conducted with each isolate by using concentrations ranging between 0.25 and 4 times the MIC(80) . The MIC(80)s for the test isolates ranged from 0.0039 to 0.25 micro g/ml . Overall, the addition of serum or plasma increased the MIC 6 to 7 doubling dilutions for C . albicans and 3 to 4 doubling dilutions for C . krusei and C . tropicalis . Micafungin time-kill studies demonstrated fungicidal activity at concentrations ranging from 4 to 16 times the MIC(80) . Micafungin is very potent agent against a variety of Candida spp., producing fungicidal activity against 7 of 10 isolates tested . A PAFE was observed against all isolates . The PAFE was influenced by the drug concentration, with the highest concentration resulting in the longest observed PAFE in each case . The highest concentration tested, four times the MIC, resulted in a PAFE of more than 9.8 h for 5 of 10 isolates tested (range, 0.9 to > or =20.1 h).

Int J Food Microbiol, 2003 Feb 15, 80(3), 187 - 99
Microbiological and fermentation characteristics of togwa, a Tanzanian fermented food; Mugula JK et al.; Selected microbiological and metabolic characteristics of sorghum, maize, millet and maize-sorghum togwa were investigated during natural fermentation for 24 h . The process was predominated by lactic acid bacteria (LAB) and yeasts . The mesophiles, lactic acid bacteria, and yeasts increased and the Enterobacteriaceae decreased to undetectable levels within 24 h . The isolated microorganisms were tentatively identified as Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus fermentum, Lactobacillus cellobiosus, Pediococcus pentosaceus, Weissella confusa, Issatchenkia orientalis, Saccharomyces cerevisiae, Candida pelliculosa and Candida tropicalis . The pH decreased from 5.24-5.52 to 3.10-3.34 . Maltose increased initially and then decreased, fructose decreased and glucose levels increased during the first 12 h of fermentation . The organic acids detected during fermentation included DL-lactic, succinic, formic, pyruvic, citric, pyroglutamic and uric acid . Lactate was the predominant acid and increased significantly with time . The volatile organic compounds (VOC) detected included acetaldehyde, 2-methyl-propanal, 2-methyl-butanal, 3-methyl-butanal, ethanol, 2-methyl-1-propanol, 2-methyl-1-butanol, 3-methyl-1-butanol, diacetyl and acetoin . Ethanol was the predominant VOC and it increased significantly with time .

J Clin Microbiol, 2002 Nov, 40(11), 4308 - 12
Rapid identification and differentiation of Candida albicans and Candida dubliniensis by capillary-based amplification and fluorescent probe hybridization; Selvarangan R et al.; We developed a rapid genotypic assay to differentiate the germ tube-positive yeasts Candida albicans and Candida dubliniensis . Fluorescently labeled nucleic acid probe binding and subsequent denaturation from the target site in the PCR amplicons produced characteristic peak melting temperatures (T(m)) that identified each species . Peak T(m)s of C . albicans (n = 69) and C . dubliniensis (n = 28) isolates produced in the presence of their respective probes were 61.04 +/- 0.64 degrees C and 60.52 +/- 1.01 degrees C (averages +/- standard deviations) . No signal was generated when the C . albicans or C . dubliniensis probes were tested against DNA from their counterparts . Both probes reacted with Candida tropicalis DNA, but the T(m) was 51.85 +/- 0.05 degrees C with the C . albicans probe and 51.92 +/- 0.10 degrees C with the C . dubliniensis probe, differentiating C . tropicalis DNA from C . albicans and C . dubliniensis . A novel hybrid probe was designed to identify both species in a single reaction based on a 4 degrees C difference in peak T(m)s . Our assay is rapid (</=2 h) and allows reliable detection and differentiation of the two germ tube-positive Candida spp.

Phytomedicine, 2002 Sep, 9(6), 566 - 71
Bioactivity of crude extracts and some constituents of Blutaparon portulacoides (Amaranthaceae); Salvador MJ et al.; Crude extracts (aerial parts and roots, both dried), methylenedioxyflavonol, and a mixture of acyl steryl glycosides isolated from Blutaparon portulacoides, were assayed for their toxicity against Trypanosoma cruzi trypomastigotes and Leishmania amazonensis amastigotes from axenic cultures . The antimicrobial activity was also investigated, in a screening conducted using fifteen strains of Gram-positive and Gram-negative bacteria, along with the yeasts, Candida albicans and Candida tropicalis . To assess the antibacterial activity of the isolated compounds, the minimum inhibitory concentrations (MICs) were determined . There are no reports of acyl steryl glycosides in the genus Blutaparon and their biological activities are being evaluated for the first time.

Antimicrob Agents Chemother, 2002 Nov, 46(11), 3532 - 9
Histone deacetylase inhibitors enhance Candida albicans sensitivity to azoles and related antifungals: correlation with reduction in CDR and ERG upregulation; Smith WL et al.; Histone acetylation and deacetylation play important roles in eukaryotic gene regulation . Several histone deacetylase (HDA) inhibitors have been characterized, including trichostatin A (TSA), apicidin, and sodium butyrate . We tested their effects on Candida albicans in vitro growth, heat sensitivity, and germ tube formation; minimal effects were observed . However, there was a dramatic effect of TSA on C . albicans sensitivity to the azoles fluconazole, itraconazole, and miconazole . Similar effects were observed with other HDA inhibitors and with the antifungals terbinafine and fenpropimorph, which target, as do azoles, enzymes in the ergosterol biosynthetic pathway . In contrast, HDA inhibitors had minimal effect on the activities of amphotericin B, flucytosine, and echinocandin, which have unrelated targets . Specifically, addition of 3 micro g of TSA/ml lowered the itraconazole MIC for five susceptible C . albicans isolates an average of 2.7-fold at 24 h, but this increased to >200-fold at 48 h . Thus, the primary effect of TSA was a reduction in azole trailing . TSA also enhanced itraconazole activity against Candida parapsilosis and Candida tropicalis but had no effect with four less related yeast species . To examine the molecular basis for these effects, we studied expression of ERG genes (encoding azole and terbinafine targets) and CDR/MDR1 genes (encoding multidrug transporters) in C . albicans cells treated with fluconazole or terbinafine with or without TSA . Both antifungals induced to various levels the expression of ERG1, ERG11, CDR1, and CDR2; addition of TSA reduced this upregulation 50 to 100% . This most likely explains the inhibition of azole and terbinafine trailing by TSA and, more generally, provides evidence that trailing is mediated by upregulation of target enzymes and multidrug transporters.

Infection, 2002 Oct, 30(5), 286 - 91
In vivo pathogenicity of eight medically relevant Candida species in an animal model; Arendrup M et al.; BACKGROUND: The relative pathogenicity of eight medically important Candida species was investigated in a mouse model . MATERIALS AND METHODS: Seventeen isolates were included; two isolates of Candida albicans, Candida glabrata, Candida tropicalis, Candida lusitaniae, Candida parapsilosis, Candida kefyr and Candida guilliermondii and three Candida krusei isolates . Mice in groups of three or four were inoculated with 10(5) and 10(7) CFU, respectively . On days 2 and 7 kidneys were removed, weighed and CFU/g kidney determined by the spot technique . RESULTS: Mortality was only observed in mice inoculated with C . albicans and C . tropicalis . Inoculated with 10(7) CFU C . tropicalis, C . glabrata, C . kefyr, C . lusitaniae, C . parapsilosis, C . krusei and C . guilliermondii the median log CFU/g kidney was significantly different: 6.0, 6.0, 6.4, 7.0, 3.7, < 2 and < 2 respectively (p < 0.0001) . Eye infection and histological changes of kidneys were examined for C . albicans, C . tropicalis, C . glabrata and C . krusei-infected mice . Weight loss, kidney weight, inflammation and infection and number of eyes infected decreased with the pathogenicity of the four species . CONCLUSION: The virulence was highly different, illustrated by a 7-log difference in CFU/g kidney, a span of 0-100% mortality and histological changes in kidneys ranging from discrete to serious . The species could be divided into three groups with decreasing virulence: 1) C . albicans and C . tropicalis, 2) C . glabrata, C . kefyr and C . lusitaniae and 3) C . parapsilosis, C . krusei and C . guilliermondii . To our knowledge this is the first study with a simultaneous comparison of all eight species in an immunocompetent animal model.

Diagn Microbiol Infect Dis, 2002 Sep, 44(1), 11 - 6
Hematogenous infections due to Candida parapsilosis: changing trends in fungemic patients at a comprehensive cancer center during the last four decades; Safdar A et al.; This study was performed to evaluate trends in species distribution in patients' with hematogenous candidiasis at a comprehensive cancer center . The results of a retrospective analysis from January 1, 1993 to December 31, 1998 were compared with prior reports from Memorial Sloan-Kettering Cancer Center in the last forty years . In 570 total episodes since 1974, 43.9% were due to Candida albicans . During 1990's, C . parapsilosis emerged as the most frequent yeast species in the non-C . albicans group (36.1% during 1993-1998 from 20.9% 1974-1982; p < 0.01) . An increase in C . krusei from 5.9% (1974-1982) to 10.5% during the recent six years (1993-1998) was also noticed . The proportion of C . tropicalis among non-albicans fungemia during 1974-1982 was 42.8%, whereas in 1993 to 1998 a marked decline in C . tropicalis hematogenous infection was observed (27.8%; p < 0.01) . During 1998, the incidence of candidemia declined from 7.1% (1972-1973) and 6.5% (1982) to 3.4% (p < 0.01), and improved survival among fungemic patients (33% mortality in 1998; 77.3% during 1974-1982; p < 0.001) was encouraging . The increase in C . parapsilosis bloodstream invasion during 1990's was associated with a significant reduction in the endogenous non-albicans Candida tropicalis infection that probably resulted in part due to the common prophylaxis, and/or preemptive fluconazole given routinely in high-risk patients undergoing treatment for cancer . The widespread use of extraneous implantable and/or semi-implantable indwelling intra-vascular devices may also have played an important role in promoting (exogenous) C . parapsilosis infection . This study emphasizes the importance of periodic evaluation of candidemia, especially at centers caring for patients at risk.

Oral Dis, 2002 Jul, 8(4), 199 - 206
Impact of lysozyme and lactoferrin on oral Candida isolates exposed to polyene antimycotics and fluconazole; Anil S et al.; OBJECTIVE: To assess the antifungal activity of lysozyme and lactoferrin on 10 oral isolates each of Candida albicans and Candida tropicalis following their brief exposure (I h) to subtherapeutic concentrations of two polyene drugs - nystatin, amphotericin B, and an azole -fluconazole . METHODS: Yeasts were sequentially exposed to subtherapeutic concentrations of antifungals and then to either lysozyme or lactoferrin and the viability evaluated by quantifying colony-forming units . RESULTS: The exposure of both C . albicans and C . tropicalis isolates to all three antifungal agents significantly increased their susceptibility to lysozyme (P < 0.05) but not to lactoferrin . Exposure to the two polyene drugs had a lesser impact on the lysozyme susceptibility of both Candida species compared with the azole drug . Both interspecies and intraspecies sensitivity to lysozyme was noted and C . albicans was less susceptible than C . tropicalis . CONCLUSIONS: Lysozyme, in addition to being a potent natural antifungal agent, may act synergistically with the studied antimycotics.

Appl Environ Microbiol, 2002 Sep, 68(9), 4517 - 22
Construction of yeast strains with high cell surface lipase activity by using novel display systems based on the Flo1p flocculation functional domain; Matsumoto T et al.; We constructed a novel cell-surface display system, using as a new type of cell-wall anchor 3,297 or 4,341 bp of the 3' region of the FLO1 gene (FS or FL gene, respectively), which encodes the flocculation functional domain of Flo1p . In this system, the N terminus of the target protein was fused to the FS or FL protein and the fusion proteins were expressed under the control of the inducible promoter UPR-ICL (5' upstream region of the isocitrate lyase of Candida tropicalis) . Using this new system, recombinant lipase with a pro sequence from Rhizopus oryzae (rProROL), which has its active site near the C terminus, was displayed on the cell surface . Cell-surface display of the FSProROL and FLProROL fusion proteins was confirmed by immunofluorescence microscopy and immunoblotting . Lipase activity reached 145 IU/liter (61.3 IU/g {dry cell weight}) on the surface of the yeast cells, which successfully catalyzed the methanolysis reaction . Using these whole-cell biocatalysts, methylesters synthesized from triglyceride and methanol reached 78.3% after 72 h of reaction . To our knowledge, this is the first example of cell-surface display of lipase with high activity . Interestingly, the yeast cells displaying the FLProROL protein showed strong flocculation, even though the glycosylphosphatidylinositol anchor attachment signal and cell-membrane-anchoring region of Flo1p had been deleted from this gene . The cell-surface display system based on FL thus endows the yeast strain with both novel enzyme display and strong flocculation ability.

J Hosp Infect, 2002 Aug, 51(4), 297 - 304
European Confederation of Medical Mycology (ECMM) prospective survey of candidaemia: report from one Italian region; Tortorano AM et al.; An ECMM epidemiological prospective survey of candidaemia was performed in one Italian region (Lombardy; population: 8 924 870) by the National Society of Medical Mycology (FIMUA) from September 1997 to December 1999 . In total, 569 episodes were reported with an overall rate of 0.38/1000 admissions, 4.4/100000 patient days . Predisposing factors included presence of an intravascular catheter (89%), antibiotic treatment (88%), surgery (56%), intensive care (45%), solid tumour (28%), steroid treatment (15%), haematological malignancy (7%), HIV infection (6%), fetal immaturity (4%) . Mucous membrane colonization preceded candidaemia in 83% of patients . Candida albicans was identified in 58% of cases, followed by Candida parapsilosis (15%), Candida glabrata (13%), Candida tropicalis (6%) . Septic shock occurred in 95 patients . Crude mortality was 35%, the highest in C . tropicalis fungaemia (44%), the elderly (64%) and solid tumour cancer patients (43%) . Intravascular catheter removal was associated with higher survival rate (71 vs . 47%) . This survey underscores the importance of candidaemia in hospital settings.

J Int Med Res, 2002 May-Jun, 30(3), 322 - 4
Esterase activity in various Candida species; Aktas E et al.; The aim of the present study is to ascertain esterase activities of various species of Candida . A total of 125 strains isolated and identified by conventional methods were tested for esterase activity using the Tween 80 opacity test . Our results showed that 58 of 59 strains of Candida albicans, all of the Candida tropicalis strains (n = 38) and all of the Candida guilliermondii strains (n = 3) produced positive results, whereas the remaining Candida species did not . The Tween 80 opacity test is a useful method because it is simple, economical and easy to prepare and interpret.

Laryngoscope, 2002 Apr, 112(4), 708 - 12
Colonization of voice prostheses by albicans and non-albicans Candida species; Bauters TG et al.; OBJECTIVES: The purposes of the study were to assess the colonization of tracheoesophageal voice prostheses by albicans and non-albicans Candida species and to determine their susceptibility for three antimycotics that are frequently used for prophylaxis or treatment of oral candidiasis (i.e., miconazole, fluconazole, and nystatin) . STUDY DESIGN: In total, 101 patients, corresponding to 170 voice prostheses, were monitored over a period of 28 months . METHODS: An enzymatic two-step method was used for differentiation and presumptive identification of Candida species colonizing the voice prostheses . The identity of the isolates was confirmed by the germ-tube test, morphological appearance on cornmeal agar with 0.5% Tween 80, sugar assimilation tests, and appearance on CHROMagar Candida (CHROMagar Co., Paris), Albicans ID (BioMerieux Vitek, Hazelwood, MO), and Fluoroplate Candida (Merck, Darmstadt, Germany) . Susceptibility testing for miconazole, fluconazole, and nystatin was performed according to the microdilution method of the National Committee for Clinical Laboratory Standards . RESULTS: The predominant species isolated were Candida albicans (41.4%), Candida glabrata (33.1%), Candida krusei (15.9%), and Candida tropicalis (5.3%) . A broad range of minimal inhibitory concentrations of the isolates was observed for miconazole and fluconazole . In contrast, minimal inhibitory concentration values for nystatin were narrowly distributed around 4 microg/mL for all isolates, suggesting uniform sensitivity . CONCLUSION: Our data on the prevalence and susceptibility of yeast isolates will contribute to a rational choice of an antimycotic for prophylaxis of the early deterioration and leakage of tracheoesophageal voice prostheses.

J Clin Microbiol, 2002 Aug, 40(8), 2860 - 5
Rapid identification of pathogenic fungi directly from cultures by using multiplex PCR; Luo G et al.; A multiplex PCR method was developed to identify simultaneously multiple fungal pathogens in a single reaction . Five sets of species-specific primers were designed from the internal transcribed spacer (ITS) regions, ITS1 and ITS2, of the rRNA gene to identify Candida albicans, Candida glabrata, Candida parapsilosis, Candida tropicalis, and Aspergillus fumigatus . Another set of previously published ITS primers, CN4 and CN5, were used to identify Cryptococcus neoformans . Three sets of primers were used in one multiplex PCR to identify three different species . Six different species of pathogenic fungi can be identified with two multiplex PCRs . Furthermore, instead of using templates of purified genomic DNA, we performed the PCR directly from yeast colonies or cultures, which simplified the procedure and precluded contamination during the extraction of DNA . A total of 242 fungal isolates were tested, representing 13 species of yeasts, four species of Aspergillus, and three zygomycetes . The multiplex PCR was tested on isolated DNA or fungal colonies, and both provided 100% sensitivity and specificity . However, DNA from only about half the molds could be amplified directly from mycelial fragments, while DNA from every yeast colony was amplified . This multiplex PCR method provides a rapid, simple, and reliable alternative to conventional methods to identify common clinical fungal isolates.

N Z Dent J, 2002 Jun, 98(432), 36 - 9
Antifungal drug susceptibilities of commensal Candida isolates; Holmes AR et al.; Two hundred and forty-two oral commensal yeast isolates were obtained from a convenience sample of 134 healthy 7- and 8-year-old children (65 males and 69 females) . The isolates were initially tested for their susceptibilities to the antifungal azole drug fluconazole, using an agar diffusion method (Etest), which was suitable for screening large numbers of yeast isolates, and confirmed as equivalent to the broth microdilution reference method . Eighteen isolates from 7 children were found to have low fluconazole susceptibility according to guidelines published by the United States National Committee for Clinical Laboratory Standards (NCCLS) . The isolates with low susceptibility were identified as either Candida tropicalis (n = 9 of 34 strains tested) or Candida glabrata (n = 9 of 13 strains tested) . Selected isolates (6 susceptible and 7 with lower susceptibility to fluconazole) were also tested by a reference broth microdilution method for susceptibility to fluconazole and to a related over-the-counter azole antifungal, miconazole . A positive correlation between susceptibility to fluconazole and to miconazole was observed . The high rate (38 percent) of reduced susceptibility in commensal C tropicalis and C glabrata strains may represent a future treatment problem if the use of over-the-counter azole drugs increase.

Appl Environ Microbiol, 2002 Jul, 68(7), 3622 - 7
New chromogenic agar medium for the identification of Candida spp; Cooke VM et al.; A new chromogenic agar medium (Candida diagnostic agar {CDA}) for differentiation of Candida spp . is described . This medium is based on Sabouraud dextrose agar (Oxoid CM41) and contains (per liter) 40.0 g of glucose, 10.0 g of mycological peptone, and 15.0 g of agar along with a novel chromogenic glucosaminidase substrate, ammonium 4-(2-{4-(2-acetamido-2-deoxy-beta-D-glucopyranosyloxy)-3-methoxyphenyl}-vinyl)-1-(propan-3-yl-oate)-quinolium bromide (0.32 g liter(-1)) . The glucosaminidase substrate in CDA was hydrolyzed by Candida albicans and Candida dubliniensis, yielding white colonies with deep-red spots on a yellow transparent background after 24 to 48 h of incubation at 37 degrees C . Colonies of Candida tropicalis and Candida kefyr were uniformly pink, and colonies of other Candida spp., including Candida glabrata and Candida parapsilosis, were white . CDA was evaluated by using 115 test strains of Candida spp . and other clinically important yeasts and was compared with two commercially available chromogenic agars (Candida ID agar {bioMerieux} and CHROMagar Candida {CHROMagar Company Ltd.}) . On all three agars, colonies of C . albicans were not distinguished from colonies of C . dubliniensis . However, for the group containing C . albicans plus C . dubliniensis, both the sensitivity and the specificity of detection when CDA was used were 100%, compared with values of 97.6 and 100%, respectively, with CHROMagar Candida and 100 and 96.8%, respectively, with Candida ID agar . In addition, for the group containing C . tropicalis plus C . kefyr, the sensitivity and specificity of detection when CDA was used were also 100%, compared with 72.7 and 98.1%, respectively, with CHROMagar Candida . Candida ID agar did not differentiate C . tropicalis and C . kefyr strains but did differentiate members of a broader group (C . tropicalis, C . kefyr, Candida lusitaniae plus Candida guilliermondii); the sensitivity and specificity of detection for members of this group were 94.7 and 93.8%, respectively . In addition to the increased sensitivity and/or specificity of Candida detection when CDA was used, differentiation of colony types on CDA (red spotted, pink, or no color) was unambiguous and did not require precise assessment of colony color.

Fungal Genet Biol, 2002 Jul, 36(2), 117 - 27
Involvement of Candida albicans NADH dehydrogenase complex I in filamentation; McDonough JA et al.; The gene encoding the 51-kDa subunit of nicotinamide adenine dinucleotide (NADH) dehydrogenase complex I, a principal component of the mitochondrial electron transport chain, was cloned in Candida tropicalis . The homolog in C . albicans, CaNDH51, was identified, and each allele was successively disrupted by PCR-mediated gene disruption . Wild type, heterozygote, reintegrant, and homozygous null mutants grew as blastoconidia in rich medium containing 3% glucose, but the homozygous null mutant failed to grow in ethanol or acetate . When glucose concentration was varied from 1 mM (0.018%) to 200 mM (3.6%) in a basal salts medium, all strains grew equally well at all glucose concentrations; the wild-type strain, the heterozygote, and the reintegrant exhibited abundant germ tubes, pseudohyphae, and hyphae . In contrast, the ndh51/ndh51 strain failed to display any type of filamentous growth, even in glucose concentrations as low as 1 mM . These results suggest a previously unexplored relationship between mitochondrial electron transport and morphogenesis.

J Ind Microbiol Biotechnol, 2002 Jul, 29(1), 16 - 9
Optimization of fed-batch fermentation for xylitol production by Candida tropicalis; Kim JH et al.; Xylitol, a functional sweetener, was produced from xylose by biological conversion using Candida tropicalis ATCC 13803 . Based on a two-substrate fermentation using glucose for cell growth and xylose for xylitol production, fed-batch fermentations were undertaken to increase the final xylitol concentration . The effects of xylose and xylitol on xylitol production rate were studied to determine the optimum concentrations for fed-batch fermentation . Xylose concentration in the medium (100 g l(-1)) and less than 200 g l(-1) total xylose plus xylitol concentration were determined as optimum for maximum xylitol production rate and xylitol yield . Increasing the concentrations of xylose and xylitol decreased the rate and yield of xylitol production and the specific cell growth rate, probably because of an increase in osmotic stress that would interfere with xylose transport, xylitol flux to secretion to cell metabolism . The feeding rate of xylose solution during the fed-batch mode of operation was determined by using the mass balance equations and kinetic parameters involved in the equations in order to increase final xylitol concentration without affecting xylitol and productivity . The optimized fed-batch fermentation resulted in 187 g l(-1) xylitol concentration, 0.75 g xylitol g xylose(-1) xylitol yield and 3.9 g xylitol l(-1) h(-1) volumetric productivity.

Boll Chim Farm, 2002 Jan-Feb, 141(1), 3 - 7
Synthesis and antimicrobial evaluation of new phenoxyacetamide derivatives; Raffa D et al.; New N-(5-methylisoxazol-3-yl)-2 or 3 or 4-(phenoxyacetamido)benzamides 6a-t were synthesized and tested for their in vitro antimicrobial activity against gram positive (Staphylococcus aureus ATCC 25923) and gram negative (Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853) bacteria as well as fungi (Candida albicans ATCC 10231, Candida tropicalis ATCC 13803 and Cryptococcus neoformans ATCC 90112) . Compounds 6 were devoid of antibacterial as well as antifungal activities at maximum tested concentrations of 50 micrograms/ml for bacteria and 100 micrograms/ml for yeast.

Diagn Microbiol Infect Dis, 2002 May, 43(1), 13 - 7
Antifungal activities of fluconazole, caspofungin (MK0991), and anidulafungin (LY 303366) alone and in combination against Candida spp . and Crytococcus neoformans via time-kill methods; Roling EE et al.; The activities of the echinocandins caspofungin and anidulafungin were evaluated alone and in combination with fluconazole using time-kill methods against isolates of Candida albicans, Candida glabrata, Candida tropicalis, Candida krusei, and Cryptococcus neoformans . Antifungal concentrations tested against each isolate were 0.5 microg/mL and 20 microg/mL of fluconazole and 0.007 microg/mL and 2 microg/mL of both caspofungin and anidulafungin . In addition, 20 microg/mL of fluconazole was tested with 2 microg/mL of caspofungin and anidulafungin to test for additive or antagonistic activity . Finally 0.5 microg/mL of fluconazole was tested with 0.007 microg/mL of caspofungin and anidulafungin to test for synergy . Combinations of fluconazole and caspofungin or anidulafungin resulted in indifference . Azole-echinocandin combinations do not produce antagonistic effects; therefore, combinations of these agents may warrant future clinical evaluation.

J Antimicrob Chemother, 2002 Jun, 49(6), 981 - 7
Antifungal susceptibilities of Candida spp . isolated from blood in Spain and Argentina, 1996-1999; Cuenca-Estrella M et al.; The aim of this study was to identify retrospectively trends in species distribution and susceptibility patterns of Candida species causing bloodstream infections in 99 medical centres (55 in Spain and 44 in Argentina) from 1996 to 1999 . A total of 744 Candida isolates were sent to the mycology reference laboratories during the study period (514 to the Spanish laboratory and 230 to the Argentinian laboratory) . Candida non-albicans strains caused more episodes of fungaemia than Candida albicans isolates in both Spain and Argentina . C . albicans was isolated in 30.2% (155/514) and 40.9% (94/230) of episodes in Spain and in Argentina, respectively . In addition, Candida parapsilosis was the second most commonly isolated pathogen (36.4%) . Candida tropicalis caused 13.7% of infections and Candida glabrata 7.4% . The amphotericin B MIC was <or=1 mg/L for 97.5% of isolates, and 8.3% of strains had decreased susceptibility to flucytosine . Regarding susceptibility to azole agents, 9.9% (74/744) and 21.9% (163/744) exhibited decreased susceptibility to fluconazole and itraconazole, respectively . For Candida species, some marked differences were found between countries, and decreased susceptibility to azole agents was detected significantly more frequently (P < 0.05) among Argentinian isolates of C . albicans, C . parapsilosis and C . tropicalis . These findings reinforced the need for continued surveillance programmes to analyse the factors that may have an influence on candidaemia incidence . Susceptibility patterns were obtained by means of the proposed reference procedure for antifungal susceptibility testing of the European Committee on Antibiotic Susceptibility Testing (EUCAST) . Excellent interlaboratory agreement was achieved for MICs for quality control strains noted in Spain and in Argentina (intraclass correlation coefficient of 0.97), indicating that the EUCAST procedure is a reliable methodology for susceptibility testing.

J Biochem (Tokyo), 2002 Jun, 131(6), 821 - 31
Structurally and functionally conserved domains in the diverse hydrophilic carboxy-terminal halves of various yeast and fungal Na+/H+ antiporters (Nha1p); Kamauchi S et al.; Genes encoding the Na(+)/H(+) antiporter (Nha1p) from Candida tropicalis (C.t.), Hansenula anomala (H.a.) (also named Pichia anomala), and Aspergillus nidulans (A.n.) were cloned, and the nucleotide sequences were determined . The deduced primary sequences revealed highly conserved hydrophobic regions and rather diverse hydrophilic regions . Among the seven known Nha1p sequences, Schizosaccharomyces pombe (S.p.) Nha1p is exceptional in lacking the hydrophilic region . Within the diverse hydrophilic regions, we found six conserved regions (C1-C6) . Expression of C.t . Nha1p in Saccharomyces cerevisiae (S.c.) cells lacking NHA1 and ENA1 (Na(+)-ATPase) complemented the salinity-sensitive phenotype, suggesting that C.t . Nha1p is functionally related to S.c . Nha1p . Expression of various truncated forms of the C-terminal half of S.c . and C.t . Nha1p showed essentially the same phenotype for both species: deletion of the C4-C6 region caused cell growth to be more resistant to high salinity than the wild type, suggesting an inhibitory function of these domains on the antiporter activity . However, complete loss of C1-C6 caused a severe growth defect under conditions of high salinity, suggesting a defect in antiporter activity . The DeltaC2-C6 form of C.t . Nha1p, containing only C1, restored the retarded cell growth at high salinity more than the control vector alone, but to a value lower than the wild type . These results suggest an essential role for C1 and an activating role of the C2-C3 region in the functional expression of Nha1 . High expression of the DeltaC2-C6 form of S.c . Nha1p was toxic for yeast cells, although low expression was not, suggesting that the overexpression of C1 is toxic . The results in this study suggest that the diverse hydrophilic region of yeast and fungal Nha1p has six conserved domains with conserved functions in terms of expression of Nha1p activity.

Indian J Pathol Microbiol, 2001 Jul, 44(3), 313 - 4
Prevalence, species distribution and antifungal sensitivity of vaginal yeasts in infertile women; Verghese S et al.; A total of 326 high vaginal swabs from infertile women attending the Institute of Reproductive Medicine were cultured from June 1999 to May 2000 . Candida species was isolated from 42(12.88%) patients . Candida albicans (40.47%) followed by Candida glabrata (38.09%) were the most common isolates . Other species included Candida tropicalis (14.28%) and Candida krusei (7.14%) . All isolates were tested for sensitivity by disc diffusion method on Yeast Nitrogen Agar base towards four antifungals . Seven (16.7%) candida strains showed resistance to Fluconazole and 19(45.23%) of the strains showed resistance to Itraconazole, and 4(9.5%) strains showed resistance to Nystatin . There was no resistance to Amphotericin B . Of the 7 strains resistant to Fluconazole, 3 were Candida krusei, 3 were Candida glabrata and 1 was Candida tropicalis.

J Hosp Infect, 2002 Apr, 50(4), 316 - 9
Candida tropicalis fungaemia in adult patients with haematological malignancies: clinical features and risk factors; Leung AY et al.; Candida tropicalis fungaemia is a serious opportunistic infection . Eighteen consecutive patients with C . tropicalis fungaemia diagnosed within a five-year period were studied retrospectively . All patients had haematological malignancies treated by chemotherapy or bone marrow transplantation (BMT) . Antifungal prophylaxis included nystatin (20 mg daily) for patients receiving chemotherapy, and fluconazole (200 mg daily) for patients undergoing BMT . Sixteen patients had refractory and advanced haematological malignancies . All patients were neutropenic, had central venous catheters, and were receiving treatment with broad-spectrum antibiotics at the time of fungaemia . Septic shock with skin emboli were the most common presenting features . In seven cases, fungaemia was preceded by a positive culture of C . tropicalis in the urine . Concomitant bacteraemia was found in 11 cases, of which six cases were due to Staphylococcus aureus . The overall mortality rate was 56% . The predominance of chemotherapy-treated patients developing fungaemia in this series might be attributable to the omission of fluconazole prophylaxis . The clinicopathologic features and risk factors identified in this study may help design better treatment strategies for this often-lethal complication .

Mycopathologia, 2002, 153(4), 195 - 8
Nosocomial candidemia in a tertiary care hospital in Saudi Arabia; Bukharie HA; Demographic information, risk factors, therapy, and outcome for all patients who had candidemia at King Fahad teaching hospital Al-khobar, between January 1995 and January 2000 were retrospectively reviewed . Thirty-two candidemic patients were identified . Candida parapsilosis was the most frequently isolated species (44%), followed by Candida tropicalis (25%), Candida albicans (19%), Candida krusei (6%), Candida glabrata (3%), and Candida guilliermondi (3%) . Risk factors included recent broad-spectrum antibiotics use (100%), ICU residency (71%), central venous catheters (66%), recent surgery (56%), total parenteral nutrition (43%), and immunosuppressive therapy (31%) . Fluconazole was used before the onset of candidemia in only two patients . The overall mortality rate was 44% . Eight (25%) episodes of candidemia were not diagnosed and treated before the patient's demise . In view of the high mortality rate associated with hematogenous candidiasis, and lack of sensitive and specific laboratory tests necessary for the premortem diagnosis of infection, empirical antifungal therapy is recommended for high-risk patients.

Mycopathologia, 2002, 153(4), 179 - 85
The impact of polyene, azole, and DNA analogue antimycotics on the cell surface hydrophobicity of Candida albicans and Candida tropicalis in HIV infection; Anil S et al.; Oral candidiasis is the most common opportunistic infection in individuals infected with the human immunodeficiency virus . Though Candida albicans is the major aetiological agent, non-albicans species such Candida tropicalis are now emerging as important agents of such infection . The Candida cell surface hydrophobicity (CSH) is considered a critical factor contributing to its colonization potential and virulence . It is also known that brief exposure to sub-cidal concentrations of antifungal agents is a likely scenario in the oral environment where the administered drugs are diluted continuously due to the flushing action of saliva . Hence the objective of the present study was to compare the CSH of 10 isolates each of C . albicans and C . tropicalis from HIV-infected individuals following brief exposure (1 hour) of isolates to sub-therapeutic concentrations of nystatin, amphotericin B, ketoconazole, fluconazole and 5-flurocytosine . The CSH was assessed by a previously described biphasic aqueous-hydrocarbon assay . The mean percentage reduction of CSH of C . albicans following brief exposure to nystatin, amphotericin B, ketoconazole, fluconazole and 5-flurocytosine was 27.33 (p < 0.001), 21.34 (p < 0.05), 11.74 (p > 0.05), 18.4 (p > 0.05) and 14.64 (p > 0.05) respectively . The mean percentage reduction of CSH of C . tropicalis following brief exposure to nystatin, amphotericin B, ketoconazole, fluconazole and 5-flurocytosine was 33.81 (p < 0.01), 28.88 (p < 0.01), 12.6 (p > 0.05), 21.53 (p > 0.05) and 17.68 (p > 0.05) respectively . A significant interspecies variation in CSH was observed for nystatin and amphoterecin B . Overall the results reveal that the CSH of C . albicans is affected to a significantly lesser degree compared with C . tropicalis when exposed to the antifungals . These data further illustrate another mode of action of antifungals on Candida leading to a reduction in the CSH and thereby the yeast adherence to host tissues.

Environ Pollut, 2002, 118(3), 379 - 82
Isolation and characterization of biarylic structure-degrading yeasts: hydroxylation potential of dibenzofuran; Romer MC et al.; Yeast communities from heavily polluted sediments that received the discharge from oil refineries and other industries were studied . Yeast species were isolated from these sediments and their ability to degrade dibenzofuran were determined . Twenty-four different yeast strains were isolated and cultured on aromatic medium; two Candida krusei strains . Candida tenuis, Candida tropicalis, two Pichia anomala strains, Pichia haplophila, two Rhodotorula glutinis strains, Rhodotorula mucilaginosa, two Trichosporon pullulans strains and Yarrowia lipolytica were able to hydroxylate dibenzofuran . Three metabolites were identified by HPLC analysis: 3-hydroxydibenzofuran was in all the cases the most abundant isomer, and while 4-hydroxydibenzofuran was also common, 2-hydroxydibenzofuran was detected in very small quantities and with few species . In the R . glutinis and Y . lipolytica cultures a ring cleavage product was also found . While in the R . gluttinis assays the hydroxydibenzofuran was detected earlier, at 2 days' incubation time, in the other yeast experiments they were observed at the 4-5th incubation days with the maximum amounts at the 7th day . Our results confirmed the ability of autochthonous yeast species to hydroxylate dibenzofuran and to cleave the rings, and it is the first report for C . krusei, C . tenuis, P . anomala, P . haplophila and R . mucilaginosa . The ecological relevance of this study is based on the fact that dibenzofuran is a xenobiotic not easily transformed, so the catabolic activities observed in authochonous yeasts contribute to broadening the biodegradable substrate spectrum.

Eur J Med Chem, 2002 May, 37(5), 409 - 18
Synthesis and preliminary evaluation of benzimidazole derivatives as antimicrobial agents; Klimesova V et al.; A series of 2-alkylsulphanylbenzimidazoles was synthesised and the compounds were evaluated for their in vitro antimicrobial activity . The structures of the compounds were confirmed by 1H-NMR and IR data, and their purity by elemental analysis . Antimycobacterial activities against Mycobacterium tuberculosis and non-tuberculous mycobacteria as well as antifungal activities against Candida albicans, Candida tropicalis, Candida krusei, Candida glabrata, Trichosporon beigelii, Trichophyton mentagrophytes and Aspergillus fumigatus were expressed as the corresponding MIC values . The substances exhibited appreciable antimycobacterial activity, in particular, against non-tuberculous mycobacteria . The activity of the most active compound in the set, 3,5-dinitro derivative 4t, exceeded that of the standard isoniazide against M . kansasii and M . avium . The antifungal activities of the compounds were relatively low . A weak antifungal effect was observed against the dermatophyte Trichophyton mentagrophytes . None of the compounds showed significant inhibitory activity against yeasts.

Appl Biochem Biotechnol, 2001 Spring, 91-93, 423 - 35
Model compound studies: influence of aeration and hemicellulosic sugars on xylitol production by Candida tropicalis; Walthers T et al.; The influence of other hemicellulosic sugars (arabinose, galactose, mannose, and glucose), oxygen limitation, and initial xylose concentration on the fermentation of xylose to xylitol was investigated using experimental design methodology . Oxygen limitation and initial xylose concentration had strong influences on xylitol production by Candida tropicalis ATCC 96745 . Under semiaerobic conditions, xylitol yield was highest (0.62 g/g), whereas under aerobic conditions volumetric productivity was highest (0.90 g/{L x h}) . In the presence of glucose, xylose utilization was strongly repressed and sequential sugar utilization was observed . Ethanol produced from the glucose caused a 50% reduction in xylitol yield when the ethanol concentration exceeded 30 g/L . When complex synthetic hemicellulosic sugars were fermented, glucose was initially consumed followed by a simultaneous uptake of the other sugars . The highest xylitol yield (0.84 g/g) and volumetric productivity (0.49 g/{L x h}) were obtained for substrates containing high arabinose and low glucose and mannose contents.

Appl Microbiol Biotechnol, 2002 Mar, 58(4), 511 - 6 Epub 2002 Feb 01.
Controlled transient changes reveal differences in metabolite production in two Candida yeasts; Granstrom T et al.; Physiological responses during growth on xylose and the xylose-degrading pathway of Candida tropicalis and Candida guilliermondii yeasts were investigated . The responses to a linearly decreasing oxygen transfer rate and a simultaneously increasing dilution rate were compared . C . guilliermondii produced acetate but no ethanol, and C . tropicalis ethanol but no acetate under oxygen limitation . Both strains produced glycerol . The D-xylose reductase of C . guilliermondii is exclusively NADPH-dependent . and acetate production regenerated NADPH . The xylose'reductase of C . tropicalis has a dual dependency for both NADH and NADPH . It regenerated NAD by producing ethanol . Both strains regenerated NAD by producing glycerol . The effect of intracellular NADH accumulation to xylose uptake and metabolite production was studied by using formate as a cosubstrate . Formate feeding in C . tropicalis triggered the accumulation of glycerol, ethanol and xylitol . Consequently, the specific xylose consumption increased 28% during formate feeding, from 477 to 609 C-mmol/C-mol cell dry-weight (CDW)/h . In C . guilliermondii cultures . formate feeding resulted only in glycerol accumulation . The specific xylose consumption increased 6%, from 301 to 319 C-mmol/C-mol CDW/h, until glycerol started to accumulate.

FEBS Lett, 1971 Nov 15, 19(1), 45 - 49

Gallo M, Bertrand JC, Azoulay E.
Candida tropicalis strain 101 possesses a hydroxylase system when grown on tetradecane as the carbon source which is active towards hydrocarbons and fatty acids . This system including cytochrome P(450) and NADPH-cytochrome c reductase has been localized exclusively in the microsomal fraction.

J Clin Microbiol, 2002 Mar, 40(3), 918 - 21
Susceptibility testing of fluconazole by the NCCLS broth macrodilution method, E-test, and disk diffusion for application in the routine laboratory; Vandenbossche I et al.; Antifungal susceptibility testing may be an important aid in the treatment of patients with life-threatening yeast infections . In order to establish the suitability of different susceptibility test methods for fluconazole with yeasts, the Rosco tablet and the E-test were compared with the gold standard NCCLS broth macrodilution method for 106 yeast strains . These included 102 clinical isolates of Candida spp., including Candida glabrata (n = 30), Candida albicans (n = 20), Candida tropicalis (n = 13), Candida parapsilosis (n = 10), Candida krusei (n = 8), plus Cryptococcus neoformans (n = 3), Saccharomyces cerevisiae (n = 2), and 16 strains belonging to other Candida spp . Four American Type Culture Collection strains of Candida were included as quality controls . The NCCLS method was found to be too complex and labor-intensive for routine testing . The E-test is an accurate alternative, but experience in determining MICs and careful attention to procedural details are critically important . The Rosco tablet showed the best agreement with the NCCLS reference method, especially when newly established breakpoints of R < or = 10 mm and S > or = 21 mm were used.

Clin Microbiol Infect, 1996 Feb, 2(3), 202 - 208
Routine use of CHROMagar Candida medium for presumptive identification of Candida yeast species and detection of mixed fungal populations; Bouchara JP et al.; OBJECTIVE: To assess the value of the new differential culture medium CHROMagar Candida for routine investigation of clinical specimens . METHODS: During a whole year, 6150 clinical samples were plated on CHROMagar Candida medium . After incubation, the green colonies were considered to be Candida albicans . The colonies of other colors were identified using Bichrolatex-krusei, or by their assimilation pattern on ID 32C test strips and their morphology on rice cream-agar-Tween . RESULTS: Among the 6150 clinical samples, 1643 were positive for fungi . Aspergillus fumigatus and Geotrichum sp . were the predominant filamentous fungi isolated . Candida albicans was the most common species isolated (1274 of the positive samples; 77.5%), and Candida glabrata was the second most common yeast isolated (174 positive samples; 10.6%) . Other yeast species were detected at lower frequencies, mainly Candida tropicalis (3.8%), Candida krusei (2.7%), Saccharomyces cerevisiae (2.7%) and Candida kefyr (2.3%), and 16 samples revealed a lipophilic species, Malassezia furfur . Mixed fungal populations accounted for 14.7% of the positive samples . Two or more yeast species were detected in 206 of the 242 specimens containing mixed fungal populations, and five yeast species were detected in one sample . Additionally, we did not observe significant differences in the isolation of yeasts or filamentous fungi from the 366 samples simultaneously plated on CHROMagar Candida and Sabouraud dextrose agar . Close agreement between the two culture media was observed for 89.9% of these samples . CONCLUSIONS: CHROMagar Candida medium was shown to be extremely helpful in a routine clinical mycology service, facilitating the detection of mixed cultures of yeasts and allowing direct identification of C . albicans, as well as rapid presumptive identification of the other yeasts: C . glabrata, C . tropicalis, C . krusei and S . cerevisiae . This chromogenic medium thus appears to be suitable as a primary culture medium, particularly for the mycologic surveillance of immunocompromised patients.

Clin Microbiol Infect, 1997 Jun, 3(3), 369 - 375
Comparison of Auxacolor with API 20 C Aux in yeast identification; Willemsen M et al.; OBJECTIVE: To compare Auxacolor with API 20 C Aux for identification of yeasts . METHODS: A total of 206 isolates belonging to 25 species was used in this study . Conventional yeast identification methods were used as a reference . RESULTS: With API 20 C Aux, the correct identification rate was 89.3% after 2 days, while 94.7% of the strains were correctly identified after 3 days . One of 14 strains of Candida tropicalis and 10 of 16 strains of Trichosporon cutaneum were not correctly identified . With Auxacolor, the percentages of correct identification after 1 and 2 days were 60.1% and 69.4%, respectively . Most strains of 11 of the 20 species considered in the system were correctly identified, including several of the most frequent yeast species . Several less commonly encountered yeast species were not correctly identified . Suggestions for improvement of the Auxacolor system are given . CONCLUSIONS: For the most frequent yeast species, Auxacolor, after adaptation and correction of the identification table, provides a useful alternative to API 20 C Aux . For less frequently encountered yeast species, the use of API 20 C Aux is preferable.

Eur J Clin Microbiol Infect Dis, 2001 Dec, 20(12), 864 - 70
Contribution of serological tests and blood culture to the early diagnosis of systemic candidiasis; Yera H et al.; The isolation of Candida species from a single blood culture is considered sufficient evidence for the initiation of systemic antifungal therapy . However, blood cultures still lack sensitivity . Previous reports have suggested that the combined serological detection of mannanemia and anti-mannan antibodies may be useful for the diagnosis of systemic candidiasis caused by Candida albicans (specificity and sensitivity 93% and 80%, respectively) . In this study, serological tests to detect Candida albicans mannan and Candida albicans antibodies (Platelia Candida Antigen and Antibody tests; Bio-Rad, France) were applied retrospectively to a series of patients with at least one Candida-positive blood culture and from whom at least one serum sample, taken before or on the day of blood culture, was available . Forty-five patients were selected, including 23 infected by Candida albicans, 4 by Candida glabrata, 9 by Candida tropicalis, 5 by Candida parapsilosis, and 4 by Candida krusei . Serological tests were positive in 73% of patients at least 2 days, and in some patients, up to 15 days before blood cultures became positive . These data suggest that serological surveillance of at-risk patients using the Platelia Candida tests could result in earlier initiation of antifungal therapy, especially when used in conjunction with blood cultures . In this way, more efficient management of nosocomial infections caused by Candida species can be achieved.

Folia Histochem Cytobiol, 2001, 39 Suppl 2, 154 - 5
Heavy metals biosorption from aqueous solution by simple eucaryotic organisms; Zielinski R et al.; Biosorption of cadmium and chromium (III) ions by means of selected yeast species has been estimated . Kinetics and equilibrium measurements have shown the reliable efficiency of both metals removal for Candida tropicalis . The influence of pH and ionic strength on biosorption process has been examined as well . For both metals the adsorption isotherms have been presented . The equilibrium of chromium (III) sorption has appeared compatible to Langmiur model and the maximum sorption capacity has been determined.

J Antimicrob Chemother, 2002 Feb, 49(2), 415 - 9
Real-time antifungal susceptibility screening aids management of invasive yeast infections in immunocompromised patients; Hadley S et al.; We examined the utility of a semi-solid agar antifungal susceptibility screening (SAAS) test in real-time management of eight immunocompromised patients with invasive yeast infections . Tests of amphotericin B and fluconazole concentrations of 0.5 and 2 mg/L and 1, 8 and 40 mg/L, respectively, were performed on Candida albicans (two), Candida tropicalis (two), Candida krusei (one), Candida glabrata (one) and Trichosporon species (spp.) (two) . All but the Trichosporon spp . and C . glabrata isolates were resistant to fluconazole at > or = 40 mg/L, and patients were successfully managed accordingly . Real-time antifungal susceptibility screening can assist in clinical management of invasive yeast infections.

Curr Genet, 2001 Dec, 40(4), 282 - 7
A transformation system for the biocontrol yeast, Candida oleophila, based on hygromycin B resistance; Yehuda H et al.; Lithium acetate transformation and electroporation were applied to the biocontrol yeast, Candida oleophila . The hygromycin B resistance gene, flanked by the phosphoglycerate kinase promoter and terminator of Candida tropicalis, served as the genetic selection marker . The transformation efficiency of electroporation was almost 400 times more efficient than that of the lithium acetate method . While incorporation of DNA, flanked by a sequence endogenous to C . oleophila, transpired apparently by homologous recombination, the integration of DNA (that did not contain C . oleophila DNA) occurred at random . Whereas transformants were observed with a linear segment of the plasmid, none were detected with the undigested plasmid . This system provides both a tool for the molecular analysis of the biocontrol mechanism of C . oleophila and a means of tagging C . oleophila for field studies.

J Antimicrob Chemother, 2002 Feb, 49 Suppl 1, 69 - 73
Neonatal candidosis: clinical picture, management controversies and consensus, and new therapeutic options; Leibovitz E; Candida infections are increasingly being recognized as a major cause of septicaemia in neonatal intensive care units, and are associated with high morbidity (25%) and mortality (25-54%) . Low birth weight pre-term infants are especially vulnerable to this devastating disease . The most frequently encountered fungal infections are caused by Candida albicans, Candida parapsilosis and, rarely, by Candida tropicalis . Amphotericin B (with or without flucytosine) is the treatment of choice for Candida infections in neonates . Conventional amphotericin B use is often limited by its severe side effects, although these tend to be fewer in neonates than in adults . Possible alternatives to amphotericin B include triazoles (such as fluconazole) and lipid preparations of amphotericin B . Liposomal encapsulation of amphotericin B has been shown to decrease the toxicity of the drug while maintaining its antifungal activity . The liposomal formulation AmBisome has proved to be effective in the treatment of severe fungal infections in adult and paediatric immunocompromised patients who fail to respond to conventional amphotericin B . The experience with AmBisome in the treatment of fungal infections in neonates is limited, and the drug has been used mainly in infants either failing conventional amphotericin B or having intolerable toxicity . Pharmacokinetic studies have not yet been performed in neonates . Three uncontrolled studies published between 1997 and 1998 on AmBisome (dose range 1-7 mg/kg/day) in the treatment of neonatal candidosis revealed that the drug was effective and safe . New information is accumulating on the safe use of high-dose AmBisome (5-7 mg/kg/day) in very low birth weight infants, and successful use of the drug as first-line therapy of neonatal candidosis . These promising results suggest a potential role for AmBisome as an additional first-line treatment of systemic candidosis in neonates.

J Antimicrob Chemother, 2002 Feb, 49 Suppl 1, 7 - 10
Amphotericin B: spectrum and resistance; Ellis D; Amphotericin B is a polyene macrolide antibiotic derived from the actinomycete Streptomyces nodosus . Of the 200 known polyene agents, amphotericin B is the only one with toxicities that are sufficiently limited to permit intravenous administration . All polyenes have a common mechanism of action in that they preferentially bind to ergosterol, the primary sterol in the fungal cell membrane . The consequence of this binding includes disruption of the osmotic integrity of the membrane, with leakage of intracellular potassium and magnesium, and also the disruption of oxidative enzymes in target cells . Amphotericin B has a relatively broad spectrum of action and is useful in treating cases of candidosis, cryptococcosis, histoplasmosis, blastomycosis, paracoccidioidomycosis, coccidioidomycosis, aspergillosis, extracutaneous sporotrichosis and mucormycosis, and some cases of hyalohyphomycosis and phaeohyphomycosis . Resistance (MIC > 2 mg/L) tends to be species-dependent and emerges uncommonly and slowly in isolates from patients treated with amphotericin B . These include some individual strains of Candida albicans, Candida tropicalis, Candida parapsilosis and Candida lusitaniae, which may acquire resistance during treatment . Some isolates of Scedosporium apiospermum, Fusarium spp . and Sporothrix schenckii also show primary resistance, whereas all strains of Scedosporium prolificans demonstrate resistance . The main problems associated with the use of conventional amphotericin B have always been due to its poor aqueous solubility and toxicity rather than antifungal resistance.

J Antimicrob Chemother, 2002 Feb, 49 Suppl 1, 3 - 6
The epidemiology of candidaemia and mould infections in Australia; Slavin MA; Australian Mycology Interest Group; A retrospective review of Candida bloodstream infections (BSI) in Australian hospitals between 1995 and 1998 was performed . Nine tertiary referral hospitals in the States of New South Wales, Victoria, South Australia and Queensland participated . Of all isolates, 56% were Candida albicans, 14% Candida parapsilosis, 13% Candida glabrata, 5% Candida krusei and 4.5% Candida tropicalis . There were significant differences in the distribution of species in different patient groups . Among surgical patients, 69% of candidaemia was due to C . albicans, whereas among medical patients the proportion was 52%, and in haematology patients 43% (P = 0.0012; Pearson chi(2)) . BSI with C . krusei were almost exclusive to haematology patients and were the second leading cause of Candida infection in that group, accounting for 27% of infections . The temporal pattern of Candida isolates also revealed a relationship between the latter years of the study and a lower likelihood of infection with C . albicans . Logistic regression showed year of the study {P = 0.032; odds ratio (OR) 0.81; 95% confidence interval (CI) 0.67-0.98} and surgery (P = 0.005; OR 2.02; 95% CI 1.2-3.1) to be significant variables . The rate of candidaemia in Australian hospitals was similar to that reported for US hospitals at 0.1-0.27 per 1000 discharges . Since April 1998, the clinical database on the internet resource Mycology Online has invited submission of clinical details from cases of invasive mycoses from Australian clinicians . To date, descriptions of 43 patients with proven or presumptive mould infection have been entered on the database . Of these, the leading cause was infection with Aspergillus (n = 16), followed by Zygomycetes (n = 7), Fusarium (n = 5), Scedosporium (n = 5) and Exserohilum (n = 1) . Although Aspergillus infections were the most frequent on the database, the variety of mould infections seen in this short time was surprising . A knowledge of local patterns of infection and antifungal susceptibility is useful in selecting empirical therapy and formulating prophylactic and pre-emptive strategies.

Zhonghua Yi Xue Za Zhi (Taipei), 2001 Oct, 64(10), 568 - 74
A comparison of methods for yeast identification including CHROMagar Candida, Vitek system YBC and a traditional biochemical method; Huang LU et al.; BACKGROUND: CHROMagar Candida (CAC) is a new chromogenic medium for the presumptive identification of clinically-important yeast isolates . A yeast biochemical card (YBC), a part of the Vitek system is an automatic method for the identification of clinically-important yeast isolates . We conducted a comparison of these two methods with a traditional biochemical method in order to choose a rapid and accurate technique for yeast identification . METHODS: All yeast isolates were inoculated onto Sabourand dextrose agar (SDA) and CAC, and incubated at 30 degrees C for 48 hours . All isolates were simultaneously tested using traditional biochemical methods and the yeast biochemical card from the Vitek system . RESULTS: We evaluated 235 yeast isolates from clinical specimens, including 89 Candida albicans, 47 Candida tropicalis, 43 Candida glabrata, six Trichosporon beigelii, and five Candida krusei in addition to 45 isolates of other yeast species . Isolates were presumptively identified on the basis of colony color and appearance on CAC medium . These observations were compared with a traditional biochemical yeast-identification method and also with YBC from the Vitek system . For five commonly-isolated species (Candida albicans, Candida tropicalis, Candida glabrata, Candida krusei and Trichosporon beigelii), agreement among the CAC medium, YBC method and traditional biochemical method were 98.9% (187/189), 96.3% (182/189), 100% (189/189), respectively . CONCLUSIONS: From our comparison, the CAC medium is a convenient and economic method to identify five commonly-noted yeast species, and the YBC method warrants a greater cost and requires a longer period of time to obtain reliable results.

Appl Microbiol Biotechnol, 2001 Dec, 57(5-6), 770 - 5
Controlled release of water-soluble polymeric complexes of sorbic acid with antifungal activities; Charvalos E et al.; We synthesized six water-soluble polymeric complexes of sorbic acid with polyvinylpyrrolidone of different molecular weight (mol wt) . As shown by infrared absorption spectrum analysis, the complexes were formed by hydrogen bonding . The complexes (SC1, with mol wt=10 kDa, SC2 with mol wt=25 kDa, SC3 with mol wt=30 kDa, SC4 with mol wt=40 kDa, SC5 with mol wt=90 kDa, and SC6 with mol wt=360 kDa) were characterized as low mol wt (SC1, SC2, and SC3) and high mol wt (SC4, SC5, and SC6) . The antifungal potencies of the complexes were tested by the macrodilution susceptibility method against environmental and clinically important fungi . Sorbic acid as well as the complexes exhibited minimum inhibitory concentrations (MICs) lower than potassium sorbate against all the strains tested . MICs of SC1, SC2, and SC3 were shown to be 2- to 4-fold lower for yeast and 1.5- to 3-fold lower than those of sorbic acid for moulds, respectively . The MICs of SC4 and SC5 against both of the Candida species tested ranged from 500 to 800 microg/ml, whereas for SC6 and sorbic acid they were about 1 mg/ml . The potencies of the high mol wt complexes against moulds were decreased by increasing the mol wt . For both of the moulds tested, the MICs of SC4 were slightly lower than those of sorbate . The MICs of sorbic acid and SC5 were equal to 300 microg/ml and 500 microg/ml respectively for Aspergillus parasiticus and for Penicillum viridicatum . The susceptibility to SC6 of all of the hyphomycetes tested was higher than that to sorbic acid . The low mol wt complexes and the sorbic acid exhibited minimal fungicidal concentrations (MFCs) 2 and 3 times higher respectively than the MICs . Sorbic acid and SC3 at a concentration of 2.5 mg/ml in an in vitro time kill curve study of Candida tropicalis were shown to be fungistatic, whereas SC1 and SC2 were fungicidal at the same concentrations . For Aspergillus parasiticus sorbic acid at 2.5 mg/ml was fungistatic for a 24-h period, whereas SC1, SC2, and SC3 were fungicidal.

Clin Rheumatol, 2001, 20(6), 435 - 7
Spondylodiscitis due to Candida tropicalis as a cause of inflammatory back pain; Sebastiani GD et al.; Spondylodiscitis may be either infectious or rheumatic in origin . In the latter case it may be seen more often in the context of spondyloarthropathies, giving rise to inflammatory back pain . We report the case of a man, affected by ulcerative colitis and carcinoma of the colon, who developed spondylodiscitis due to infection by Candida tropicalis.

Mycoses, 2001 Nov, 44(9-10), 375 - 8
Antifungal activity of propolis on different species of Candida; Ota C et al.; Propolis is a resinous material collected by bees from the buds or other parts of plants . It is known for its biological properties, having antibacterial, antifungal and healing properties . The antifungal activity of propolis was studied in sensitivity tests on 80 strains of Candida yeasts: 20 strains of Candida albicans, 20 strains of Candida tropicalis, 20 strains of Candida krusei and 15 strains of Candida guilliermondii . The yeasts showed a clear antifungal activity with the following order of sensitivity: C . albicans > C . tropicalis > C . krusei > C . guilliermondii . Patients with full dentures who used a hydroalcoholic propolis extract showed a decrease in the number of Candida.

Appl Microbiol Biotechnol, 2001 Nov, 57(4), 528 - 33
Creation of cell surface-engineered yeast that display different fluorescent proteins in response to the glucose concentration; Shibasaki S et al.; We have successfully created a novel yeast strain able to monitor changes in environmental conditions by displaying either green fluorescent protein (GFP) from Aequorea victoria or blue fluorescent protein (BFP), a variant of GFP, on its cell surface as a visible reporter . For the display of these fluorescent proteins on the cell surface of Saccharomyces cerevisiase, our cell-surface-engineering system was utilized . The GAPDH promoter, which is active in the presence of glucose, and the UPR-ICL promoter from Candida tropicalis, which starts to function in the presence of a reduced level of glucose, were employed simultaneously to express the GFP-encoding gene and the BFP-encoding gene, respectively . This cell-surface-engineered yeast strain emitted green fluorescence from the cell surface when sufficient glucose was present in the medium, and blue fluorescence from the same cell surface when the glucose in the medium was consumed . The fluorescent proteins displayed on the cell surface using the different promoters enabled us to monitor the concentrations of intra- and/or extracellular glucose that regulated activation or inactivation of the promoters . This novel yeast strain could facilitate the computerized control of various bioprocesses measuring emitted fluorescence.

Appl Microbiol Biotechnol, 2001 Nov, 57(4), 515 - 20
Yeast whole-cell biocatalyst constructed by intracellular overproduction of Rhizopus oryzae lipase is applicable to biodiesel fuel production; Matsumoto T et al.; Yeast whole-cell biocatalysts for lipase-catalyzed reactions were constructed by intracellularly overproducing Rhizopus oryzae lipase (ROL) in Saccharomvces cerevisiae MT8-1 . The gene encoding lipase from R . orvzae IFO4697 was cloned, and intracellular overproduction systems of a recombinant ROL with a pro-sequence (rProROL) were constructed . When rProROL from R . oryzae IFO4697 was produced under the control of the 5'-upstream region of the isocitrate lyase gene of Candida tropicalis (UPR-ICL) at 30 degrees C for 98 h by two-stage cultivation using SDC medium (SD medium with 2% casamino acids) containing 2.0% and 0.5% glucose, intracellular lipase activity reached levels up to 474.5 IU/l . These whole-cell biocatalysts were permeabilized by air-drying and used for the synthesis of methyl esters (MEs), a potential biodiesel fuel, from plant oil and methanol in a solvent-free and water-containing system . The ME content in the reaction mixture was 71 wt% after a 165-h reaction at 37 degrres C with stepwise addition of methanol . These results indicate that an efficient whole-cell biocatalyst can be prepared by intracellular overproduction of lipase in yeast cells and their permeabilization.

Appl Microbiol Biotechnol, 2001 Nov, 57(4), 500 - 5
Development of novel whole-cell immunoadsorbents by yeast surface display of the IgG-binding domain; Nakamura Y et al.; The ZZ domain derived from Staphylococcus aureus, which binds to the Fc part of immunoglobulin G (IgG), was displayed on the cell surfaces of yeast Saccharomyces cerevisiae by cell-surface engineering using the C-terminal half of alpha-agglutinin under control of the 5'-upstream region of the isocitrate lyase gene from Candida tropicalis (UPR-ICL) . Display of ZZ on the cell surface was confirmed by immunofluorescence microscopy . Enzyme-linked immunosorbent assay (ELISA) and sandwich ELISA using the S . cerevisiae cells displaying ZZ detected IgG and antigen (human serum albumin) down to a concentration of 1-10 ng/ml in both cases . The detection range covered by these assay systems was wide and could be varied by adjusting the amount of cells and reaction times with horseradish peroxidase (HRP) substrate . Moreover, yeast cells displaying ZZ were successfully used for repeated affinity purification of IgG from serum . These results indicate that S . cerevisiae displaying ZZ may constitute novel and genetically renewable whole-cell immunoadsorbents widely applicable to immunoassays and affinity purification.

Bioorg Med Chem Lett, 2002 Jan 21, 12(2), 117 - 20
Antifungal sordarins . Synthesis and structure-activity relationships of 3'-O-substituted derivatives; Arribas EM et al.; A number of novel 3'-O-acyl and alkyl sordarins were synthesised for structure-activity relationship studies . Many of these derivatives exhibit high activity against Candida albicans, Candida pseudotropicalis, Candida tropicalis and Cryptococcus neoformans.

Yeast, 2001 Nov, 18(15), 1365 - 70
A nuclear-encoded intein in the fungal pathogen Cryptococcus neoformans; Butler MI et al.; We have used comparative sequence analysis to identify an intein-like sequence (protein splicing element) present in Cryptococcus neoformans, a fungal pathogen of humans . The sequence encoding this element is present in the C . neoformans PRP8 gene, as an in-frame insertion relative to the PRP8 genes of other organisms . It contains sequences similar to those of the protein-splicing domains of two previously described yeast inteins (in Saccharomyces cerevisiae and Candida tropicalis), although it lacks any recognizable internal endonuclease domain . The Cryptococcus neoformans intein (Cne PRP8) is only the second to be found in a eukaryote nuclear genome; the previously described yeast inteins occur at the same site in the VMA gene homologues of S . cerevisiae and C . tropicalis . The host gene of the Cryptococcus intein, PRP8, encodes a highly conserved mRNA splicing protein found as part of the spliceosome . The Cne PRP8 intein may be a useful drug target in addressing the cryptococcal infections so prevalent in AIDS patients .

Saudi Med J, 2001 Oct, 22(10), 860 - 3
In vitro activity of 6 antifungal agents on candida species isolated as causative agents from vaginal and other clinical specimens; Ellabib MS et al.; OBJECTIVE: To study the susceptibility pattern of candida species isolated from various clinical specimens to common usable antifungals in Libya . METHODS: Two hundred and four candida species were isolated from patients complaining of fungal infections gathered from vaginal swabs, nails, throat, hair and ear . Yeast isolates were identified to the species level by API 20C AUX Commercial system . The in vitro susceptibility to amphotericin B, nystatin, ketoconazole, miconazole, clotrimazole and econazole was determined using the macrodilution in broth method . RESULTS: Candida albicans was the most common isolated species from vaginal and Candida tropicalis from throat swabs . On the other hand Candida parapsilosis and Candida guilliermondii were more often isolated from the ear and hair . Candida krusei and Candida fomata were only isolated from vulvaginitis in this study . In vitro sensitivity showed that most of the isolates were inhibited at optimum ranges of minimal inhibitory concentration particularly with amphotericin B and nyastatin . On the other hand resistance strains of candida species were found against the 4 azoles antifungal agents . CONCLUSION: Candida albicans and Torulopsis glabrata are among the most common cause of vaginitis among Libyan females . Amphotericin B, nystatin and clotrimazole were the most effective antifungals against most isolates in this study . Fungicidal effect was obtained with most antifungals at higher concentrations.

FEMS Microbiol Lett, 2001 Nov 13, 204(2), 375 - 9
Physiological difference during ethanol fermentation between calcium alginate-immobilized Candida tropicalis and Saccharomyces cerevisiae; Jamai L et al.; Calcium alginate-immobilized Candida tropicalis and Saccharomyces cerevisiae are compared for glucose fermentation . Immobilized C . tropicalis cells showed a slight morphological alteration during ethanol production at 40 degrees C, but their fermentation capacity was reduced by 25% . Under immobilization conditions, the two species demonstrated two different mathematical patterns when the relationship between growth rate, respiration rate, and ethanol tolerance was assessed . The interspecific difference in behavior of immobilized yeast cells is mainly due to their natural metabolic preference . The production of CO(2) by calcium alginate-immobilized C . tropicalis, as well as the lower supply of oxygen to the cells, are the major factors that reduce ethanol production.

J Clin Microbiol, 2001 Dec, 39(12), 4387 - 9
Evaluation of Etest method for determining caspofungin (MK-0991) susceptibilities of 726 clinical isolates of Candida species; Pfaller MA et al.; The performance of the Etest for testing the susceptibilities to caspofungin (MK-0991) of 726 isolates of Candida spp . was assessed against the National Committee for Clinical Laboratory Standards (NCCLS) microdilution broth method . The NCCLS method employed RPMI 1640 broth medium, and MICs were read after incubation for 48 h at 35 degrees C . MICs were determined by Etest for all 726 isolates with RPMI agar containing 2% glucose (RPG) and were read after incubation for 48 h at 35 degrees C . The Candida isolates included Candida albicans (n = 486), Candida glabrata (n = 96), Candida tropicalis (n = 51), Candida parapsilosis (n = 47), Candida krusei (n = 11), Candida lusitaniae (n = 2), and Candida guilliermondii (n = 33) . In addition, a subset of 314 isolates were also tested by Etest using Casitone agar (CAS) and antibiotic medium 3 agar (AM3) . The Etest results obtained using RPG correlated well with reference MICs . Overall agreement was 94% with RPG, 82% with CAS, and 79% with AM3 . When RPG was used, agreement ranged from 79% for C . parapsilosis to 100% for C . krusei, C . lusitaniae, and C . guilliermondii . When CAS was used, agreement ranged from 0% for C . lusitaniae to 100% for C . glabrata . With AM3, agreement ranged from 0% for C . lusitaniae to 100% for C . guilliermondii . All three media supported growth of each of the Candida species . Etest results were easy to read, with sharp zones of inhibition . In most instances (75%) where a discrepancy was observed between the Etest and the reference method, the Etest MIC was lower . The Etest method using RPG appears to be useful for determining caspofungin susceptibilities of Candida species.

Mycoses, 2001, 44(7-8), 278 - 80
Emergence of Candida tropicalis as the major cause of fungaemia in India; Mathews MS et al.; The fungal isolates (n = 191) from 1970 consecutive blood cultures over a 9-year period were identified . Candida tropicalis was the predominant isolate accounting for 66% of all isolates whereas Candida albicans ranked only a distant second with 21% . The difference was highly significant (P < 0.00001) The other isolates included C . parapsilosis (5.8%), C . glabrata (2.1%), C . pelliculosa (1.2%), C . lipolytica (1.0%) C . krusei (1.0%) and Trichosporon beigelii (2.6%) . The isolates were obtained predominantly from postoperative patients on therapy with more than one antibiotic.

Kansenshogaku Zasshi, 2001 Oct, 75(10), 863 - 9
{An epidemiological study for fungus isolation during the twenty-five year periods from 1976 to 2000 in Kitasato University Hospital}; Obata S et al.; We investigated an epidemiological study for fungus isolation in our hospital from 1976 to 2000 . For 25 years, the total sample number of fungus examination were 64,296, and after 1988, the total sample number increased suddenly . As a whole, the positive ratio was constantly about 40% . When our hospital opened, the obstetrical and gynecological samples showed 38.8% for fungus examination, but recently, samples of the respiratory organ has increased . Ratio of isolation for yeast, Candida albicans was 53.8%, and another yeasts such as Candida glabrata, Candida tropicalis, Candida parapsilosis were 12.5%, 5.3%, and 3.4%, respectively . Recently, isolation of Candida glabrata showed a tendency to increase . For genus Aspergillus, Aspergillus fumigatus was isolated, 48.1%, and Aspergillus nigar, Aspergillus terreus were isolated, 31.4% and 7.5%, respectively . For dermatophytes, Trichophyton rubrum was isolated, 63.6% indermatophytes, and another dermatophytes were Microsporum canis (17.9%), and Trichophyton mentagrophytes (15.9%), respectively . For dermatophytes, isolation of Microsporum canis showed a tendency to increase . Recently, the plural number of species showed a tendency to increase in the samples . Compared with the number of samples at the beginning in our hospital, the plural number of species in the samples increased about six times.

Nippon Ishinkin Gakkai Zasshi, 2001, 42(4), 243 - 51
{Subtractive gene cloning using dynabeads oligo(dT)25 for elucidation of pseudohyphal formation in Candida tropicalis}; Imanishi Y et al.; The dimorphic transition from yeast to pseudohyphae in Candida tropicalis occurs following the addition of ethanol to a synthetic medium containing glucose . We developed a method of subtractive gene cloning to isolate genes, of which the expression was apparently specific for pseudohyphal formation in this organism . Subtraction was performed between sense-strand cDNAs instead of mRNAs from cells of the ethanol culture and anti-sense cDNAs linking to Dynabeads oligo(dT)25 from those of the control culture . Dynabeads oligo(dT)25 are paramagnetic beads with 25 nucleotide-long chains of deoxythymidines covalently linked to their surface and were expected to be easily collected using a magnet . This method using Dynabeads oligo(dT)25 minimizes the degradation of mRNA and makes it easy to construct a cDNA library sufficient to analyze the genetic information on the yeast-to-hyphae transition . Using this strategy, we identified several genes including a homologue of CPP1 coding tyrosine phosphatase and a homologue of nmt1+ encoding protein, which was reported to regulate thiamine biosynthesis.

Arch Microbiol, 2001 Nov, 176(5), 364 - 9
Novel and convenient methods for Candida tropicalis gene disruption using a mutated hygromycin B resistance gene; Hara A et al.; We established a novel and convenient method to construct a ura3 strain (ura3/ura3) of the asporogenous and diploid yeast, Candida tropicalis, that produces dicarboxylic acid . One copy of the URA3 gene was disrupted using a mutated hygromycin B resistance gene (HYG#) . The obtained hygromycin-resistant strain was further transformed with a URA3 disruption cassette and selected on a plate containing 5-fluoroorotic acid . The obtained strains were analyzed and the disruption of the gene was confirmed by PCR and Southern blot analysis . The results showed that the strains were obtained in which allelic URA3 genes were simultaneously disrupted . Furthermore, we established a cotransformation method for this gene disruption, using HYG# in C . tropicalis . In order to disrupt the allelic POX4 genes (encoding acyl-CoA oxidase) of dicarboxylic acid-producing strains, the ARS plasmid (which contained HYG#) and a POX4 disruption cassette (which carried the LAC4 gene encoding beta-galactosidase of Kluyveromyces lactis) were simultaneously introduced by transformation . As a result, the allelic POX4 gene was successfully disrupted.

Biotechnol Bioeng, 2001 Nov 20, 75(4), 456 - 62
Effects and mechanisms of H(2)O(2) on production of dicarboxylic acid; Jiao P et al.; The system of producing long chain dicarboxylic acid (DCA) by Candida tropicalis is an aerobic and viscous fermentation system . A method to overcome the gas-liquid transport resistance and to increase oxygen supply is by adding hydrogen peroxide (H(2)O(2)) to the fermentation system . Here we report that the H(2)O(2) not only can enhance the oxygen supply but also change the metabolism by inducing cytochrome P450, the key enzyme of a, o-oxidation . When C . tropicalis was cultivated in a 3-L bioreactor using the combination of aeration and H(2)O(2) feeding, DCA production rates increased by about 10% after a short period of decrease at the beginning . Furthermore, the experiments showed that the maximum activities of P450 could be induced at 2 mM H(2)O(2), and the inducible mechanisms are also discussed . Moreover, we suggest that alkane might be oxidized through the "peroxide shunt pathway" when H(2)O(2) is present . By adding H(2)O(2), the DCA yield in a 22-L bioreactor could increase by 25.3% and reach 153.9 g/L .

Antimicrob Agents Chemother, 2001 Nov, 45(11), 3216 - 9
Susceptibilities of oral bacteria and yeast to mammalian cathelicidins; Guthmiller JM et al.; The effects of cathelicidins against oral bacteria and clinically important oral yeasts are not known . We tested the susceptibilities of Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Porphyromonas gingivalis, Streptococcus sanguis, Candida krusei, Candida tropicalis and Candida albicans to the following cathelicidins: FALL39, SMAP29, and CAP18 . SMAP29 and CAP18 were antimicrobial, whereas FALL39 did not exhibit antimicrobial activity . Future studies are needed to determine the potential use of these antimicrobial peptides in prevention and treatment of oral infections.

Acta Trop, 2001 Oct 22, 80(2), 151 - 4
Efficacy of fluconazole and itraconazole in the treatment of oral candidiasis in HIV patients; Menon T et al.; A total of 46 strains of Candida were collected from HIV infected patients, of which 25 strains were isolated from patients with oral candidiasis, and 21 strains were from mouthwash samples of asymptomatic carriers . The most common species isolated was Candida albicans (73.9%), followed by Candida tropicalis (21.7%) . In vitro susceptibility of the strains to fluconazole and itraconazole was tested using minimum inhibitory concentration (MIC) studies by agar dilution technique . Out of the 18 strains of C . albicans isolated from mouthwash samples, four were resistant to fluconazole whereas only two were resistant to itraconazole . Out of 16 strains of C . albicans isolated from oral lesions, one was resistant to fluconazole where as all were sensitive to itraconazole . Among the other species of Candida tested, C . tropicalis gave higher MIC values to both drugs than other species such as Candida guillermondii and Candida krusei . In vitro MIC values correlated well with in vivo responses in patients . Hence, itraconazole may be used as an alternative in the treatment of candidiasis, which does not respond to fluconazole therapy.

Clin Infect Dis, 2001 Nov 15, 33(10), 1676 - 81 Epub 2001 Sep 24.
Risk Factors for Candida tropicalis fungemia in patients with cancer; Kontoyiannis DP et al.; The risk factors for and presentation of Candida tropicalis fungemia, in comparison with those of Candida albicans, have been incompletely characterized . We compared 43 cases of C . tropicalis fungemia with 148 cases of C . albicans fungemia . In univariate analysis, patients with C . tropicalis fungemia were more likely to have leukemia (P=.0006), prolonged neutropenia (P=.03), and a positive blood culture for more days (P=.02) . The 2 groups did not differ with regard to baseline Acute Physiology and Chronic Health Evaluation (APACHE) II score, frequency of catheter-associated fungemia, or response to antifungals . In multivariate analysis, patients with C . tropicalis fungemia were more likely to have leukemia (P=.02), previous neutropenia (P=.002), and a longer stay in the intensive care unit during the infectious episode (P=.01) . Also, the response of the breakthrough C . tropicalis fungemia was lower (P=.05) . In conclusion, the host determinants associated with susceptibility to C . tropicalis are leukemia and prolonged neutropenia.

Appl Microbiol Biotechnol, 2001 Aug, 56(3-4), 478 - 85
Repression of fatty-acyl-CoA oxidase-encoding gene expression is not necessarily a determinant of high-level production of dicarboxylic acids in industrial dicarboxylic-acid-producing Candida tropicalis; Hara A et al.; The synthesis of dicarboxylic acids (DCAs) in Candida tropicalis is thought to be induced by a decrease in fatty acyl-CoA-oxidase activity . However, in the present study we demonstrate that repression of the POX4 gene, encoding fatty acyl-CoA oxidase, does not directly lead to high-level production of DCAs . No fatty acyl-CoA-oxidase activity was detected if the POX4 gene of C . tropicalis strain 1098 (wild-type strain) was disrupted . Furthermore, introduction of the POX4 gene from C . tropicalis strain M1210A3, which is a mutant derived from strain 1098 and is used as an industrial DCA-producing strain, still exhibited low-level fatty acyl-CoA-oxidase activity . Nevertheless, production of DCA was not observed in either case . Furthermore, the increase in acyl-CoA-oxidase activity by expression of the POX4 gene in strain M1210A3 did not reduce high-level production of DCA . These results suggest that alterations in acyl-CoA-oxidase activity are not necessarily related to production of DCA in industrial DCA-producing C . tropicalis M1210A3.

J Oral Pathol Med, 2001 Sep, 30(8), 481 - 8
Post-antifungal effect of polyene, azole and DNA-analogue agents against oral Candida albicans and Candida tropicalis isolates in HIV disease; Anil S et al.; Oropharyngeal candidiasis (OPC) is the most frequent AIDS-associated opportunistic infection, as up to 90% of HIV-infected individuals suffer at least one episode during the course of their disease . Various in vivo and in vitro procedures have been used to assess the effectiveness of antifungal agents used in HIV infection . In the present study, we evaluated in vitro the minimum inhibitory concentration (MIC) and the post-antifungal effect (PAFE) of two polyenes, two azoles and one DNA-analogue against 10 oral isolates of Candida albicans and 10 of Candida tropicalis, all from HIV-infected individuals, in order to obtain basic data on the pharmacodynamics of these drugs . One-hour exposure to twice the MIC of all the drugs, except fluconazole, elicited a consistently high PAFE in both Candida species . Furthermore, the PAFE elicited by the antifungals (except fluconazole) was significantly prolonged for C . tropicalis compared with C . albicans . This speedy recovery of C . albicans isolates exposed to transient low concentrations of antifungals appeared to reflect its virulence compared with lesser potent species, such as C . tropicalis . Taken together, the current data, while confirming the existence of PAFE in a non-albicans species of Candida, also provide further clues for the recalcitrance of C . albicans species in the face of antifungal therapy for oropharyngeal candidiasis.

J Clin Microbiol, 2001 Sep, 39(9), 3254 - 9
International surveillance of bloodstream infections due to Candida species: frequency of occurrence and in vitro susceptibilities to fluconazole, ravuconazole, and voriconazole of isolates collected from 1997 through 1999 in the SENTRY antimicrobial surveillance program; Pfaller MA et al.; A surveillance program (SENTRY) of bloodstream infections (BSI) in the United States, Canada, Latin America, and Europe from 1997 through 1999 detected 1,184 episodes of candidemia in 71 medical centers (32 in the United States, 23 in Europe, 9 in Latin America, and 7 in Canada) . Overall, 55% of the yeast BSIs were due to Candida albicans, followed by Candida glabrata and Candida parapsilosis (15%), Candida tropicalis (9%), and miscellaneous Candida spp . (6%) . In the United States, 45% of candidemias were due to non-C . albicans species . C . glabrata (21%) was the most common non-C . albicans species in the United States, and the proportion of non-C . albicans BSIs was highest in Latin America (55%) . C . albicans accounted for 60% of BSI in Canada and 58% in Europe . C . parapsilosis was the most common non-C . albicans species in Latin America (25%), Canada (16%), and Europe (17%) . Isolates of C . albicans, C . parapsilosis, and C . tropicalis were all highly susceptible to fluconazole (97 to 100% at < or =8 microg/ml) . Likewise, 97 to 100% of these species were inhibited by < or =1 microg/ml of ravuconazole (concentration at which 50% were inhibited {MIC(50)}, 0.007 to 0.03 microg/ml) or voriconazole (MIC(50), 0.007 to 0.06 microg/ml) . Both ravuconazole and voriconazole were significantly more active than fluconazole against C . glabrata (MIC(90)s of 0.5 to 1.0 microg/ml versus 16 to 32 microg/ml, respectively) . A trend of increased susceptibility of C . glabrata to fluconazole was noted over the three-year period . The percentage of C . glabrata isolates susceptible to fluconazole increased from 48% in 1997 to 84% in 1999, and MIC(50)s decreased from 16 to 4 microg/ml . A similar trend was documented in both the Americas (57 to 84% susceptible) and Europe (22 to 80% susceptible) . Some geographic differences in susceptibility to triazole were observed with Canadian isolates generally more susceptible than isolates from the United States and Europe . These observations suggest susceptibility patterns and trends among yeast isolates from BSI and raise additional questions that can be answered only by continued surveillance and clinical investigations of the type reported here (SENTRY Program).

Haematologica . 2001 Aug;86(8):E18.
Successful treatment of hepatosplenic candidiasis in an elderly patient with acute myeloid leukemia using liposomal daunorubicin and fluconazole; Quintini G et al.; Fungal infections are an increasing cause of morbidity and mortality in patients with haematological malignancies . The organism most often responsible are Candida spp., particurarly Candida Albicans . This report describes our experience in a 63-year-old man who developed symptoms of hepatosplenic candidiasis caused by Candida tropicalis after treatment for acute myeloid leukaemia (AML) . The fungal infection was successfully controlled using fluconazole, and the patient has been disease-free for more than 11 months after antileukemic chemotherapy without any recurrence of Candida infections . Our experience suggests that AML and chemotherapy associated fungal infections can be controlled with an appropriate therapeutic regimen.

Int J Antimicrob Agents, 2001 Aug, 18(2), 179 - 83
Subtyping and antifungal susceptibilities of Candida spp . in the intensive care unit of a Greek general hospital; Kanellopoulou M et al.; This study identified the Candida spp., susceptibility to antifungal agents and the prevailing Candida albicans subtypes responsible for infections or colonization of 42 patients in the ICU over a 6-month period . Most isolates were C . albicans (66.1%) and Candida tropicalis (28.3%) all of which were susceptible in vitro to antifungal agents . Subtypes of the C . albicans isolates were identified by pulsed field gel electrophoresis Sfi I chromosomal digests . Two major C . albicans subtypes were identified, whereas subtype heterogeneity was found among strains of Candida glabrata and C . tropicalis . Sfi I PFGE restriction patterns were able to discriminate between sub-populations of C . albicans isolates, clustering them into distinct, epidemiologically congruous groups.

Mol Cell Biol, 2001 Sep, 21(18), 6243 - 53
Candida tropicalis Etr1p and Saccharomyces cerevisiae Ybr026p (Mrf1'p), 2-enoyl thioester reductases essential for mitochondrial respiratory competence; Torkko JM et al.; We report here on the identification and characterization of novel 2-enoyl thioester reductases of fatty acid metabolism, Etr1p from Candida tropicalis and its homolog Ybr026p (Mrf1'p) from Saccharomyces cerevisiae . Overexpression of these proteins in S . cerevisiae led to the development of significantly enlarged mitochondria, whereas deletion of the S . cerevisiae YBR026c gene resulted in rudimentary mitochondria with decreased contents of cytochromes and a respiration-deficient phenotype . Immunolocalization and in vivo targeting experiments showed these proteins to be predominantly mitochondrial . Mitochondrial targeting was essential for complementation of the mutant phenotype, since targeting of the reductases to other subcellular locations failed to reestablish respiratory growth . The mutant phenotype was also complemented by a mitochondrially targeted FabI protein from Escherichia coli . FabI represents a nonhomologous 2-enoyl-acyl carrier protein reductase that participates in the last step of the type II fatty acid synthesis . This indicated that 2-enoyl thioester reductase activity was critical for the mitochondrial function . We conclude that Etr1p and Ybr026p are novel 2-enoyl thioester reductases required for respiration and the maintenance of the mitochondrial compartment, putatively acting in mitochondrial synthesis of fatty acids.

Folia Microbiol (Praha), 2000, 45(6), 561 - 5
Relative pathogenicity of Candida tropicalis in rat tongue mucosa; Dorko E et al.; The potential of C . tropicalis to colonize and infect rat tongue mucosa was demonstrated . Thirty female Sprague-Dawley rats were infected orally with three different strains of C . tropicalis . The animals were killed one and three weeks following the inoculation and sections of their tongue were stained with hematoxylin-eosin and Grocott stain . Histological changes were observed in the group of animals killed one week after inoculation and infected with C . tropicalis strain isolated from the crural ulcer of a diabetic patient . The most important finding was acute purulent myositis with the formation of abscesses . The myositis was local without signs of spreading to the surrounding tissue . Epithelium-penetrating hyphae observed in the Grocott-stained sections were relatively fewer and more sparsely distributed.

Mycoses, 2001, 44(5), 147 - 9
Yeast infection of burns; Mousa HA et al.; In a prospective study, 132 patients were investigated for yeast infection of burn wounds . Ten patients (7.6%) were infected with Candida species . All patients with yeast infections were also infected with bacteria with the exception of one patient who was infected with Candida tropicalis alone . The predominant yeast recovered was Candida krusei . Yeast infection was found to be more common in the younger age group . The isolation of a Candida species alone from one patient and Candida isolation from patients with sepsis in burn wounds indicate a significant role for yeasts in the production of infection in burn wounds . Therefore, special cultures for yeasts are recommended for all cases of burn wound infection.

J Antimicrob Chemother, 2001 Aug, 48(2), 245 - 51
Treatment of experimental candidosis with amphotericin B-Intralipid admixtures in immunocompromised mice; Shadkchan Y et al.; The main goal of this research was the evaluation of the efficacy of amphotericin B (AMB) in comparison with AMB-Intralipid (AMB-IL) admixtures in cyclophosphamide (CY)-compromised animals for the treatment of systemic candidosis induced by several pathogenic Candida spp . Four-week-old ICR female mice were inoculated ip with 200 mg/kg of CY . At day 4 post-CY treatment the animals were inoculated iv with Candida albicans, Candida glabrata or Candida tropicalis (different inocula for the different species) . Forty-eight hours later various doses of conventional AMB (0.4-1 mg/kg for 5 days) or AMB-IL admixtures (0.4-2 mg/kg for 5 days) were administered iv and the survival rate and mean survival time (MST) were evaluated during an observation period of up to 42 days . These experiments showed that while all control animals died, the survival rate of the AMB-treated mice ranged between 13 and 65% and that of the AMB-IL-treated mice was in the range 30-100% depending on the infecting dose and Candida species . The follow-up of the course of infection showed that AMB-IL admixtures increased the survival time of the treated mice . The MST was significantly higher for the mice treated with AMB-IL than for those treated with conventional AMB and was especially marked in the groups treated with high doses of the drug . Hence, the data obtained in the present study show that in CY-compromised mice AMB-IL admixtures were very effective in the treatment of systemic candidosis caused by C . albicans and non-albicans species.

J Clin Microbiol, 2001 Aug, 39(8), 2933 - 6
Evaluation of BACTEC MYCO/F Lytic medium for recovery of mycobacteria, fungi, and bacteria from blood; Fuller DD et al.; MYCO/F Lytic medium (MFL), a liquid medium developed for use with the BACTEC 9240 blood culture system, was compared to the Isolator system (IS) for the recovery of fungi and to the BACTEC 13A medium for the recovery of mycobacteria . Recovery of bacteria was compared to routine BACTEC Plus Aerobic/F (AF) blood cultures . Microbial growth was detected in 203 (17%) of 1,166 blood cultures . Fifty-seven specimens were positive for fungi: 35 were positive with both IS and MFL; six were positive with IS only (three Candida albicans, one Histoplasma capsulatum, one Candida glabrata, and one Fusarium species isolate); three were positive with AF only (two C . albicans and one Candida parapsilosis isolate); and 13 were positive with MFL only (five C . glabrata, three C . albicans, two Candida krusei, two Candida tropicalis, and one C . parapsilosis isolate; P > 0.05 versus IS) . Eighteen of 19 blood cultures positive for H . capsulatum grew in both IS and MFL, although the time to detection for MFL was greater . The mean time to detection for all fungi was 8.15 days for IS and 12.07 days for MFL . Seven hundred forty specimens were also cultured for mycobacteria with MFL and 13A . Forty-four grew mycobacteria; 38 were positive with both 13A and MFL; and 16 were positive with MFL only . Mycobacterium avium was recovered from 41 specimens; 36 were positive for both systems and 5 were positive for MFL alone . MFL was also compared to the AF bottle for the same 740 specimens . MFL and AF both detected 34 of the 40 clinically significant bacteria, while IS detected only 15 of 40 . In summary, MFL is an excellent medium for the recovery of fungi, mycobacteria, and bacteria; however, the time to detection of H . capsulatum is increased.

Clin Infect Dis, 2001 Aug 15, 33(4), 523 - 30 Epub 2001 Jul 20.
Vertebral osteomyelitis due to Candida species: case report and literature review; Miller DJ et al.; Candida species uncommonly cause vertebral osteomyelitis . We present a case of lumbar vertebral osteomyelitis caused by Candida albicans and review 59 cases of candidal vertebral osteomyelitis reported in the literature . The mean age was 50 years, and the lower thoracic or lumbar spine was involved in 95% of patients . Eighty-three percent of patients had back pain for >1 month, 32% presented with fever, and 19% had neurological deficits . The erythrocyte sedimentation rate was elevated in 87% of patients, and blood culture yielded Candida species for 51% . C . albicans was responsible for 62% of cases, Candida tropicalis for 19%, and Candida glabrata for 14% . Risk factors for candidal vertebral osteomyelitis were the presence of a central venous catheter, antibiotic use, immunosuppression, and injection drug use . Medical and surgical therapies were both used, and amphotericin B was the primary antifungal agent . Prognosis was good, with an overall clinical cure rate of 85%.

Yeast, 2001 Jul, 18(10), 971 - 80
Molecular cloning of the RPS0 gene from Candida tropicalis; Baquero C et al.; The Saccharomyces cerevisiae RPS0 A and B genes encode proteins essential for maturation of the 40S ribosomal subunit precursors . We have isolated a homologue of the RPS0 gene from Candida tropicalis, which we named CtRPS0 . The C . tropicalis RPS0 encodes a protein of 261 amino acid residues with a predicted molecular weight of 28.65 kDa and an isoelectric point of 4.79 . CtRps0p displays significant amino acid sequence homology with Rps0p from C . albicans, S . cerevisiae, Neurospora crassa, Schizosaccharomyces pombe, Pneumocystis carinii and higher organisms, such as human, mouse and rat . CtRPS0 on a high copy number vector can complement the lethal phenotype linked to the disruption of both RPS0 genes in S . cerevisiae . Southern blot analysis suggests that CtRPS0 is present at a single locus within the C . tropicalis genome .

Oral Microbiol Immunol, 2001 Aug, 16(4), 250 - 2
Antifungal activity of histatin-5 against non-albicans Candida species; Nikawa H et al.; Fungicidal effects of histatin-5 against 26 oral isolates belonging to 5 non-albicans Candida species were examined . Fifty microM of histatin-5 killed more than 95% of Candida tropicalis and Candida guilliermondii isolates and more than 90% of Candida parapsilosis and Candida krusei . However, Candida glabrata was less sensitive to the peptide (mean 62.9%) . Our results, taken together, demonstrated that histatin-5 possessed the fungicidal activity against Candida species other than C . glabrata.

Infection, 2001 May-Jun, 29(3), 177 - 9
Prosthetic valve endocarditis due to Candida tropicalis complicated by multiple pseudoaneurysms; Zedtwitz-Liebenstein K et al.; Candida endocarditis is an unusual but severe complication caused by Candida albicans or other fungal species . We describe a case of prosthetic valve endocarditis due to Candida tropicalis, complicated by multiple pseudoaneurysms.

J Clin Microbiol, 2001 Jul, 39(7), 2708 - 12
Novel fluorescent broth microdilution method for fluconazole susceptibility testing of Candida albicans; Liao RS et al.; A comparative evaluation of the reference National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution method with a novel fluorescent carboxyfluorescein diacetate (CFDA)-modified microdilution method for the susceptibility testing of fluconazole was conducted with 68 Candida strains, including 53 Candida albicans, 5 Candida tropicalis, 5 Candida glabrata, and 5 Candida parapsilosis strains . We found trailing endpoints and discordant fluconazole MICs of < 8 microg/ml at 24 h and of > or =64 microg/ml at 48 h for 12 of the C . albicans strains . These strains satisfy the definition of the low-high MIC phenotype . All 12 low-high phenotype strains were correctly shown to be susceptible at 48 h with the CFDA-modified microdilution method . For the 41 non-low-high phenotype C . albicans strains, the CFDA-modified microdilution method yielded 97.6% (40 of 41 strains) agreement within +/-1 dilution at 24 h compared with the reference method and 92.7% (38 of 41 strains) agreement within +/-1 dilution at 48 h compared with the reference method . The five strains each from C . tropicalis, C . glabrata, and C . parapsilosis that were tested showed 100% agreement within +/-2 dilutions for the two methods being evaluated.

J Clin Microbiol, 2001 Jul, 39(7), 2431 - 8
Identification of two different 14-alpha sterol demethylase-related genes (cyp51A and cyp51B) in Aspergillus fumigatus and other Aspergillus species; Mellado E et al.; Two cyp51-related genes (cyp51A and cyp51B) encoding 14-alpha sterol demethylase-like enzymes were identified in the opportunistic human pathogen Aspergillus fumigatus . PCR amplification using degenerate oligonucleotides based on conserved areas of cytochrome P450 demethylases of other filamentous fungi and yeasts allowed the cloning and sequencing of two different homologue genes in A . fumigatus . Southern analysis confirmed that both genes hybridized to distinct genomic loci and that both are represented as single copies in the genome . Comparison of the deduced Cyp51A and Cyp51B proteins with the CYP51 proteins from Penicillium italicum, Aspergillus nidulans, Erysiphe graminis, Uncinula necator, Botrytis cinerea, Ustilago maydis, Cryptococcus neoformans, Candida albicans, Saccharomyces cerevisiae, Candida tropicalis, and Candida glabrata showed that the percentages of identity of the amino acid sequences (range, 40 to 70%) were high enough to consider Cyp51A and Cyp51B to be members of the fungal CYP51 family . Fragments from both genes were also cloned from other Aspergillus spp . (A . flavus, A . nidulans, and A . terreus) . Phylogenetic analysis showed that, at least in the most pathogenic species of Aspergillus, there are two fungal CYP51 proteins . This is the first report of the existence of two homologue genes coding for 14-alpha sterol demethylase in the fungal kingdom . This finding could provide insights into the azole resistance mechanisms operating in fungi . The primers used here may be useful molecular tools for facilitating the cloning of novel 14-alpha sterol demethylase genes in other filamentous fungi.

Glycoconj J, 2000 Oct, 17(10), 677 - 80
Conjugation of yeast mannans with protein employing cyanopyridinium agent (CDAP)--an effective route of antifungal vaccine preparation; Bystricky S et al.; The possibility of using 1-cyano-4-dimethylaminopyridinium tetrafluoroborate (CDAP) for activation of saccharide hydroxyl groups (instead of hazardous cyanogen bromide) is examined with cell-surface mannans of the yeasts Candida albicans, Candida tropicalis, Candida lambica and galactoglucoxylomannan of Cryptococcus laurentii . Direct conjugation with human serum albumin yielded soluble products with increased molecular size in comparison with the original polysaccharides . Immunodiffusion experiments revealed that conjugation did not affect the immunospecificity of the antigen epitppe.

Environ Technol, 2001 May, 22(5), 533 - 42
Study on sludge expansion during treatment of salad oil manufacturing wastewater by yeast; Zheng S et al.; Five yeast strains, namely Rhodotorula rubra, Candida tropicalis, Candida utilis, Candida boidinii, Trichosporon cutaneum, were isolated from soil spots of a salad oil factory, and applied for continuous treatment of salad oil manufacturing wastewater . The oil and COD removal performance of the mixed cultures were comparable to the results other researchers obtained . Sludge expansion, accompanied with sludge morphology change from pseudomycelia to true mycelia, occurred during continuous treatment of wastewater . The true mycelia dominated sludge had a much higher water content and SVI value than that of the yeast pure cultures, although the two kinds of sludge had similar oil removal performance . A mold, Geotrichum candidum, was isolated from the expanded sludge, and was suspected to be a reason for sludge expansion . Addition of 0.3% sodium propionate into batch cultures degraded SVI value from around 100 to 60 . In a continuous running, addition of 10 mg l-1 sodium hypochlorite decreased SVI value from over 200 to below 100 . The yeast activity, however, was weakened to a large extent at the same time.

Sheng Wu Gong Cheng Xue Bao, 2001 Mar, 17(2), 165 - 9
{Construction of yeast fusants of directly transform starch into ethanol}; Pang XY et al.; To obtain strains that are able to efficiently produce ethanol directly from starchy material, we have used the protoplast fusion technique to construct hybrids between Saccharomyces cerevisiae and Candida tropicalis . The isolation of fusants used a selective system by a irreversible biochemical inhibitor-Iodoacetic Acid(ID), and the fusion frequency was 9.2 x 10(-6) . The cell size and DNA content of the fusants were determined . Through testing some productive properties such as specific growth rate, saccharification ability, ethanol tolerance and the activities of alpha-amylase, saccharogen amylase, we selected two fusants--F-1 and F-5 . They were more superior and their ethanol production came up to 8.8% and 11.5%.

Eur J Clin Microbiol Infect Dis, 2001 Apr, 20(4), 276 - 9
Flucytosine primary resistance in Candida species and Cryptococcus neoformans; Cuenca-Estrella M et al.; The in vitro activity of flucytosine (5FC) against 1,140 clinical isolates of Candida spp . and Cryptococcus neoformans was evaluated and compared with the activity of amphotericin B, fluconazole and itraconazole . Overall, 87.72% (1,000/1,140) of yeasts were susceptible to 5FC . This agent showed less potent in vitro activity against Candida glabrata, Candida krusei, Candida guilliermondii and Cryptococcus neoformans (MIC90s, 8-16 microg/ml) and intermediate activity or resistance to 6.5% of Candida albicans, 5.1% of Candida tropicalis and 0.8% of Candida parapsilosis strains . Amphotericin B showed potent activity against isolates with an MIC of 5FC > or = 8 microg/ml . A total of 112 of 140 strains that were SFC-intermediate or -resistant showed decreased susceptibility to azoles (P < 0.01).

APMIS, 2001 Feb, 109(2), 108 - 12
In vitro Candidacidal effect of polymorphonuclear neutrophils of Behçet's patients with or without ocular involvement; Tekeli A et al.; In this study our objective was to evaluate the in vitro candidacidal effect of polymorphonuclear neutrophils of Behcet's patients with or without ocular involvement (inactive phase) . Fifteen patients with ocular involvement and 15 patients without ocular involvement were studied . Candidacidal assay was performed to assess the in vitro killing of Candida species by the polymorphonuclear neutrophils of both the study and control groups . In brief, peripheral venous polymorphonuclear neutrophils of healthy volunteers and patients with Behcet's disease were obtained by density gradient centrifugation . These cells were incubated with Candida spp . (Candida albicans, Candida tropicalis, Candida glabrata) for 3 h . Samples were seeded onto yeast extract peptone dextrose agar, incubated at 25 degrees C in a 5% humidified incubator for another 48 h, and formed colonies were counted . There was no significant difference between the candidacidal activity of polymorphonuclear neutrophils of healthy volunteers and of Behcet's patients with or without ocular involvement . Although various immunological abnormalities are reported in Behcet's disease, in vitro killing of Candida spp., which is one of the parameters for the phagocytic function of polymorphonuclear neutrophils, was not affected.

Mycoses, 2001, 44(1-2), 47 - 53
The utility of serology in diagnosing candidosis in non-neutropenic critically ill patients; Ibanez-Nolla J et al.; This study was carried out to evaluate the utility of serological tests in the diagnosis of candidal infections in non-neutropenic critically ill patients . A prospective study was carried out in a 10-bed general intensive care unit; all patients with at least one organic sample with Candida spp . were included . A therapeutic-diagnostic algorithm was designed, and patients were treated or not, according to a classification . Blood samples were taken, and serological tests included: antigenaemia detection using two different commercial latex kits (Cand-Tec and Pastorex) and antibody detection by indirect haemagglutination (IHA) and indirect germ tube immunofluorescence (IFA) . A total of 56% of antibody tests (IHA 45% and IFA 64%) and 26% of antigen tests (Cand-Tec 36% and Pastorex 17%) were positive . The sensitivity and specificity of these tests with respect to systemic candidosis were 37% and 78%, respectively, for antibodies, and 0% and 90% for antigens . There was statistical significance for mortality and low levels of antibodies; Candida glabrata was detected by IFA and Candida tropicalis by Cand-Tec . Serological tests may help to define the prognosis of these patients and to support the detection of specific Candida species.

Mycoses, 2001, 44(1-2), 37 - 45
Impact of endpoint definition on the outcome of antifungal susceptibility tests with Candida species: 24- versus 48-h incubation and 50 versus 80% reduction in growth; St-Germain G; The growth inhibition patterns of 764 clinical yeast isolates, in response to amphotericin B, flucytosine, itraconazole and fluconazole, were studied in order to determine the frequency of trailing growth and any impact this, as well as 24- or 48-h incubation periods, may have on minimum inhibitory concentration (MIC) results . A broth microdilution method following National Committee for Clinical Laboratory Standard No . M27A recommendations was used . Trailing growth was observed mainly with azoles . Furthermore, over 98% of isolates exhibiting a trailing effect at 24 h with fluconazole and itraconazole were either Candida albicans or Candida tropicalis . When comparing 24- and 48-h IC50 values, discrepancies were observed with itraconazole and fluconazole, respectively, in 18 and 11% of C . albicans and 24 and 30% of C . tropicalis . When comparing IC50 and IC80 values at 24 h, discrepancies were again essentially seen with itraconazole and fluconazole, respectively, in 11 and 10% of C . albicans, and 17 and 27% of C . tropicalis . In susceptibility tests performed with a microdilution method and read spectrophotometrically, 48-h IC80 values result in an unlikely high number of resistant isolates, indicating that a 24-h incubation and a 50% reduction in optical density may correlate better with clinical outcome.

Antimicrob Agents Chemother, 2001 Jun, 45(6), 1905 - 7
Azasordarins: susceptibility of fluconazole-susceptible and fluconazole-resistant clinical isolates of Candida spp . to GW 471558; Cuenca-Estrella M et al.; The in vitro activity of the azasordarin GW 471558 was compared with those of amphotericin B, flucytosine, itraconazole, and ketoconazole against 177 clinical isolates of Candida spp . GW 471558 showed potent activity against Candida albicans, Candida glabrata, and Candida tropicalis, even against isolates with decreased susceptibility to azoles . Candida krusei, Candida parapsilosis, Candida lusitaniae, and Candida guilliermondii are resistant to GW 471558 in vitro (MICs, >128 microg/ml).

Cell Biochem Biophys, 2000, 32 Spring, 285 - 90
Genetic evaluation of peroxisomal and cytosolic acetoacetyl-CoA thiolase isozymes in n-alkane-assimilating diploid yeast, Candida tropicalis; Ueda M et al.; The n-alkane-assimilating diploid yeast, Candida tropicalis, possesses two acetoacetyl-CoA thiolase (Thiolase I) isozymes encoded by one allele: peroxisomal and cytosolic Thiolase Is encoded by both CT-T1A and CT-T1B . To clarify the function of peroxisomal and cytosolic Thiolase Is, the site-directed mutation leading Thiolase I delta C6 without a putative C-terminal peroxisomal targeting signal was introduced on CT-T1A locus in the ct-t1b delta-null mutant . The C-terminus-truncated Thiolase I was active and solely present in the cytosol . Although the ct-t1a delta/t1b delta-null mutants showed mevalonate auxotrophy, the mutants having the C-terminus-truncated Thiolase I did not require mevalonate for growth, as did the strains having cytosolic Thiolase I . These results demonstrate that the presence of Thiolase I in the cytoplasm is indispensable for the sterol synthesis in this yeast . It is of greater interest that peroxisomal and cytosolic Thiolase I isozymes, products of the same genes, play different roles in the respective compartments, although further investigations will be necessary to analyze how to be sorted into peroxisomes and the cytosol.

Cell Biochem Biophys, 2000, 32 Spring, 139 - 46
Sorting of peroxisomal and mitochondrial carnitine acetyltransferase isozymes in the diploid yeast, Candida tropicalis; Tanaka A et al.; Peroxisomal and mitochondrial carnitine acetyltransferase (CAT; EC 2.3.1.7) isozymes are synthesized from the first and second ATG codons of the open reading frame of one gene, Candida tropicalis CAT . Primer extension analysis and RNase protection assay revealed that the peroxisomal CAT, initiating at the second AUG codon of the transcripts, was synthesized by a translational readthrough of the first AUG codon of the open reading frame . When C . tropicalis CAT was introduced into the other yeast, Saccharomyces cerevisiae, 5' ends of transcripts were similar to those observed in C . tropicalis . Peroxisomal and mitochondrial CAT isozymes were strongly suggested to occur by the alternative initiation of translation, chiefly dependent on the structure or sequence context of the region from the 5' end to the second AUG codon; their transcripts harbored sufficient information to bring about alternative initiation of translation in both yeasts . Sorting of peroxisomal and mitochondrial CAT isozymes to their own compartment was carried out by their own targeting sequences, but, before transportation to their destination, their biosyntheses were regulated by alternative initiation of translation.

Yeast, 2001 May, 18(7), 605 - 10
Identification of Candida tropicalis HSR1, a gene of the heat-shock factor-related family, which confers salt tolerance in Saccharomyces cerevisiae; Ali R et al.; A genomic library of Candida tropicalis in a yeast multicopy plasmid has been screened for clones conferring salt tolerance upon transformation into S . cerevisiae . The best halotolerance clone contained an open reading frame encoding a predicted protein of 728 amino acids with homology to transcription factors of the heat-shock family . This novel gene was named HSR1 and is present in single copy in the C . tropicalis genome . Upon transformation into S . cerevisiae it increases the expression of ENA1, a major determinant of salt tolerance encoding a cation-extrusion pump . The sequence of CtHSR1 has been deposited in the EMBL data library under Accession No . AJ296093 .

Eur J Biochem, 2001 May, 268(9), 2669 - 77
Secreted aspartic proteases of Candida albicans, Candida tropicalis, Candida parapsilosis and Candida lusitaniae . Inhibition with peptidomimetic inhibitors; Pichova I et al.; The frequency of Candida infections has increased in recent years and it has been accompanied by a significant rise in morbidity and mortality . The secretion of aspartic proteases by Candida spp . was demonstrated to be one of the virulence determinants . Candida albicans is classified as the major human pathogen in the genus Candida . However, other species of this genus have been found to cause an increasing number of candidiases . We isolated secreted aspartic proteases (Saps) of C . albicans (Sap2p), C . tropicalis (Sapt1p), C . parapsilosis (Sapp1p), and C . lusitaniae (Saplp) from culture media . All the isolated proteases were N-terminally sequenced . Their specific proteolytic activities and sensitivity to series of peptidomimetic inhibitors modified in the type of scissile bond replacement as well as in the N- and C-termini were analyzed . The most divergent substrate specificity was observed for the Sap of C . tropicalis . The specificity of Sap of C . lusitaniae is most closely related to that of Sap of C . parapsilosis . We designed and prepared an inhibitor containing phenylstatine isoster that was equipotent towards all four proteases within the range of 10-10-10-9 M . The HIV-1 protease inhibitors ritonavir, saquinavir, indinavir, and nelfinavir were also tested for the inhibition of four Saps . Only ritonavir and saquinavir inhibited Sap2p, Sapt1p, Sapp1p, and Saplp in micromolar concentrations.

Appl Environ Microbiol, 2001 May, 67(5), 2083 - 7
Genetically controlled self-aggregation of cell-surface-engineered yeast responding to glucose concentration; Zou W et al.; We constructed an arming (cell-surface-engineered) yeast displaying two types of agglutinin (modified a-agglutinin and alpha-agglutinin) on the cell surface, with agglutination being independent of both mating type and pheromones . The modified a-agglutinin was artificially prepared by the fusion of the genes encoding Aga1p and Aga2p . The modified a-agglutinin could induce agglutination of cells displaying Agalpha1p (alpha-agglutinin) . The upstream region of the isocitrate lyase gene of Candida tropicalis (UPR-ICL), active at a low glucose concentration, was used as the promoter to express the modified a-agglutinin- and alpha-agglutinin-encoding genes . The arming yeast displaying both agglutinins agglutinated and sedimented in response to decreased glucose concentration . When the glucose concentration was high, the arming yeast grew normally . In the late log phase, when the glucose concentration became very low, agglutination occurred suddenly and drastically and yeast cells sedimented completely . Sedimentation was confirmed by weighing the aggregated cells after filtration of the broth . Strains in which aggregation can be genetically controlled can be used in industrial processes in which the separation of yeast cells from the supernatant is necessary.

Eur J Clin Microbiol Infect Dis, 2001 Feb, 20(2), 132 - 5
Candida tropicalis and Penicillium marneffei mixed fungaemia in a patient with Waldenström's macroglobulinaemia; Wong SS et al.; A patient with Waldenstrom's macroglobulinaemia who had previously been treated with cladribine presented with septic arthritis caused by Escherichia coli . The patient's condition subsequently deteriorated, and he succumbed to pneumonia and mixed fungaemia due to Candida tropicalis and Penicillium marneffei . Profound lymphopenia coexisted with fungaemia . This is the first reported case of mixed Penicillium marneffei and Candida fungaemia and penicilliosis marneffei in a patient with Waldenstrom's macroglobulinaemia . Penicilliosis marneffei should be considered as a potential complication in patients with markedly impaired cell-mediated immunity who have travelled to or resided in endemic areas . Patients who have undergone therapy with purine nucleoside analogues such as fludarabine and cladribine are also at risk.

Antimicrob Agents Chemother, 2001 May, 45(5), 1367 - 73
Anticandida activity is retained in P-113, a 12-amino-acid fragment of histatin 5; Rothstein DM et al.; Through the analysis of a series of 25 peptides composed of various portions of the histatin 5 sequence, we have identified P-113, a 12-amino-acid fragment of histatin 5, as the smallest fragment that retains anticandidal activity comparable to that of the parent compound . Amidation of the P-113 C terminus increased the anticandidal activity of P-113 approximately twofold . The three histidine residues could be exchanged for three hydrophobic residues, with the fragment retaining anticandidal activity . However, the change of two or more of the five basic (lysine and arginine) residues to uncharged residues resulted in a substantial loss of anticandidal activity . A synthetic D-amino-acid analogue, P-113D, was as active against Candida albicans as the L-amino-acid form . In vitro MIC tests in low-ionic-strength medium showed that P-113 has potent activity against Candida albicans, Candida glabrata, Candida parapsilosis, and Candida tropicalis . These results identify P-113 as a potential antimicrobial agent in the treatment of oral candidiasis.

J Clin Microbiol, 2001 Apr, 39(4), 1422 - 8
Fluconazole and voriconazole multidisk testing of Candida species for disk test calibration and MIC estimation; Kronvall G et al.; Fluconazole and voriconazole MICs were determined for 114 clinical Candida isolates, including isolates of Candida albicans, Candida glabrata, Candida krusei, Candida lusitaniae, Candida parapsilosis, and Candida tropicalis . All strains were susceptible to voriconazole, and most strains were also susceptible to fluconazole, with the exception of C . glabrata and C . krusei, the latter being fully fluconazole resistant . Single-strain regression analysis (SRA) was applied to 54 strains, including American Type Culture Collection reference strains . The regression lines obtained were markedly different for the different Candida species . Using an MIC limit of susceptibility to fluconazole of < or =8 microg/ml, according to NCCLS standards, the zone breakpoint for susceptibility for the 25-microg fluconazole disk was calculated to be > or =18 mm for C . albicans and > or =22 mm for C . glabrata and C . krusei . SRA results for voriconazole were used to estimate an optimal disk content according to rational criteria . A 5-microg disk content of voriconazole gave measurable zones for a tentative resistance limit of 4 microg/ml, whereas a 2.5-microg disk gave zones at the same MIC level for only three of the species . A novel SRA modification, multidisk testing, was also applied to the two major species, C . albicans and C . glabrata, and the MIC estimates were compared with the true MICs for the isolates . There was a significant correlation between the two measurements . Our results show that disk diffusion methods might be useful for azole testing of Candida isolates . The method can be calibrated using SRA . Multidisk testing gives direct estimations of the MICs for the isolates.

J Clin Microbiol, 2001 Apr, 39(4), 1328 - 33
Evaluation of Etest for direct antifungal susceptibility testing of yeasts in positive blood cultures; Chang HC et al.; The performance of the Etest (AB BIODISK, Solna, Sweden) for direct antifungal susceptibility testing of yeasts in positive blood cultures was compared with that of the macrodilution method for determining the MICs of five antifungal agents . Culture broths with blood from bottles positive for yeasts were inoculated directly onto plates for susceptibility testing with the Etest, and the MICs were read after 24 and 48 h of incubation . A total of 141 positive blood cultures (72 cultures of Candida albicans, 31 of Candida tropicalis, 14 of Candida glabrata, 11 of Candida parapsilosis, 3 of Candida krusei, and 3 of Cryptococcus neoformans, 4 miscellaneous yeast species, and 3 mixed cultures) were tested, and the rates of MIC agreement (+/-1 log(2) dilution) between the direct Etest (at 24 and 48 h, respectively) and macrodilution methods were as follows: amphotericin B, 81.8 and 93.5%; flucytosine, 84.8 and 87.7%; fluconazole, 89.4 and 85.5%; itraconazole, 69.7 and 63.8%; ketoconazole, 87.9 and 79.0% . By a large-sample t test, the difference in log(2) dilution between the direct Etest and the macrodilution method was found to be small (P < 0.05) . The lone exceptions were ketoconazole at 48 h of incubation and itraconazole at both 24 and 48 h of incubation (P > 0.05) . By Tukey's multiple comparisons, the difference between the direct Etest (48 h) and reference methods among different species was found to be less than 1 log(2) dilution . When the MICs were translated into interpretive susceptibility, the minor errors caused by the direct Etest (at 24 and 48 h, respectively) were as follows: flucytosine, 2.3 and 1.4%; fluconazole, 3.0 and 3.6%; itraconazole, 21.2 and 21.3% . Itraconazole also produced an additional 3.0 and 3.6% major errors as determined by the direct Etest at 24 and 48 h, respectively . It was concluded that, except for itraconazole, the Etest method was feasible for direct susceptibility testing of blood cultures positive for yeasts . The method is simple, and the results could be read between 24 and 48 h after direct inoculation, whenever the inhibition zones were discernible.

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