Diagnostic and clinical characteristics of mycotic vaginitis caused by Candida glabrata (Torulopsis glabrata)

In this study an evaluation of frequency of occurrence of C. glabrata, its diagnosis, sensitivity to antifungal drugs and its significance in pathogenesis of mycotic vaginosis was performed. Strains belonging to C. glabrata genus constituted 12.1% of total of 852 isolated strains and 39.2% of strains other than C. albicans. During fungal vaginosis caused by C. glabrata Lactobacillus sp. was present and normal pH values of vaginal secretion were seen. In direct preparations single or few leukocytes were observed and usually numerous blastospores were present. During evaluation of the sensitivity of C. glabrata strains to antimycotic agents a decreased sensitivity of these strains to clotrimazole and ketoconazole was found what speaks for their low usefulness in the treatment of mycotic vaginosis. Significance of C. glabrata in pathogenesis of mycotic vaginosis is not questioned since release of complaints and clinical symptoms in patients with positive therapeutic effect is seen and their persistence in a group of patients with treatment failure.     

In vitro antifungal resistance in Candida albicans from HIV-infected patients with and without oral candidosis.

The main purpose of this study has been to determine the in vitro antifungal susceptibility of clinical isolates from HIV-infected or AIDS patients, depending on the presence of oral candidosis. The oral cavity of 307 HIV-infected or AIDS patients was examined and an oral swab was cultured on Sabouraud glucose agar and studied by conventional mycological methods. In vitro antifungal susceptibility to amphotericin B, nystatin, fluconazole, itraconazole and ketoconazole was tested by disk diffusion with Neo-Sensitabs tablets (Rosco Diagnostica, Dinamarca). One hundred and thirty five Candida albicans isolates (91 serotype A, 38 serotype B, three C. albicans variety stellatoidea and three untyped isolates), three Candida krusei and two Candida glabrata were obtained. All the isolates were susceptible to nystatin and amphotericin B. However, 7.9% isolates were resistant to fluconazole and 2.9% isolates were resistant to ketoconazole or itraconazole. Nearly all C. krusei and C. glabrata isolates, 31% patients with candidosis and 20% Candida-colonized patients showed decreased susceptibility to azoles. This study shows that polyenes had a great in vitro efficacy against clinical isolates from HIV-infected patients and that in vitro resistance to azoles is not as high as observed in other countries.     

Fluconazole and itraconazole susceptibility of vaginal yeast isolates from Slovakia

Vulvovaginal candidiasis is a common mucosal infection caused by opportunistic yeasts of the Candida genus. In this study, we isolated and identified the yeast species in the vagina of patients treated in the gynecology clinic and tested in vitro activities of fluconazole and itraconazole against 227 clinical yeast isolates by the NCCLS microdilution method. C. albicans (87.6%) was the most frequently identified species followed by C. glabrata (6.2%) and C. krusei (2.2%). Almost thirteen percent of yeast strains were resistant to fluconazole and 18.5% were resistant to itraconazole. Cross-resistance analyses of C. albicans isolates revealed that fluconazole resistance and itraconazole resistance were also associated with decreased susceptibilities to other azole derivatives mainly to ketoconazole and miconazole. At the same time no cross-resistance to polyene antibiotics amphotericin B and nystatin was observed. These results support the notion that antifungal agents used to treat vaginitis may be contributing to the drug resistance problem by promoting cross-resistance to a range of clinically used antifungals.     

In vitro resistance to fluconazole and itraconazole in clinical isolates of Candida spp and Cryptococcus neoformans

An in vitro susceptibility testing of 181 strains of six species of Candida and 21 strains of Cryptococcus neoformans was carried out in order to investigate the resistance to new antifungal drugs. We have studied clinical isolates from 200 different patients of Hospital del Mar (Barcelona) and Hospital La Inmaculada (Almería). An agar diffusion method (NeoSensitabs, Rosco, Taastrup, Denmark), was employed with fluconazole, itraconazole, and reference drugs amphotericin B, flucytosine, tioconazole and ketoconazole. A high level of susceptibility was found for amphotericin B in C. neoformans strains while 19% of them were resistant to flucytosine. All the strains of C. neoformans and Candida guilliermondii were susceptible to the new azoles derivatives and also Candida parapsilosis and Candida albicans had a great susceptibility to this antifungals. A greater level of resistance was found for Candida krusei, Candida tropicalis and Candida glabrata to fluconazole, itraconazole and ketoconazole, but resistance to fluconazole and itraconazole is not always linked because ten resistant strains for fluconazole were susceptible to itraconazole, and two other resistant to itraconazole were susceptible to fluconazole.     

Antifungal drug resistance pattern of Candida. spp isolated from vaginitis in Ilam-Iran during 2013-2014

Vaginal Candidiasis is the most common and important opportunistic fungal infection in women. By increasing use of antifungal drugs in recent years, it has caused drug resistance. This study aims to evaluate antifungal drugs susceptibility of Candida. spp isolated of women with vaginitis from Ilam-Iran during 2013-2014. samples were collected and cultured from 385 women with vaginitis, then Candida.spp was diagnosed by standard method. Antifungal drug susceptibility test for nystatin 100 unit/disk, fluconazole 10µg/disk, itraconazole 10µg/disk, ketoconazole 10µg/disk, amphotericinB 20µg/disk, clotrimazole 10µg/disk, posaconazole 5µg/disk, and voriconazole 1µg/disk were carried out by M44-A method(CLSI). From all culture positive samples, 150 isolates were Candida albicans and 89 isolates were non-albicans. The resistance to fluconazole, itraconazole, ketoconazole, clotrimazole, voriconazole, posaconazole, nystatin and amphotericin B was 76%, 62%, 72%, 55%, 6%, 7%, 1% and 0%. The highest resistance was seen for fluconazole , itraconazole, and the highest susceptible was seen for nystatin and amphotericin B. These results indicate nystatin and amphotericin B can be used as the first line for empirical therapy of vaginal candidiasis in the district.     

Sensitivity of fungi of the genus Candida to antifungal drugs

Seventy two Candida strains isolated from patients with candidiosis of the oral mucosa were studied with respect to their sensitivity to nystatin, levorin, decamine, ethonium, sanguiritrin and clotrimazole. At concentrations of 0.5 to 5 micrograms/ml all the Candida species i.e. C. albicans, C. tropicalis, C. krusei and C. quilliermondii were highly sensitive to clotrimazole. Fungistatic action of levorin, nystatin and sanguiritrin was observed in 91, 67 and 38 per cent of the strains respectively. The Candida strains were resistant to decamine and ethonium used in the above concentrations.     

复发性外阴阴道念珠菌病的致病菌种鉴定及药敏分析

目的：探究复发性外阴阴道念珠菌病致病菌的菌种分布及耐药情况,指导临床治疗。方法：收集210例念珠菌性外阴阴道炎患者阴道分泌物标本,其中复发性外阴阴道念珠菌病（RVVC）组76例,外阴阴道念珠菌病（VVC）组134例。运用科玛嘉显色培养基法进行菌种鉴定和药物敏感试验。结果：在210例病例中,RVVC组中白色念珠菌54株（71.05%）,光滑念珠菌15株（19.74%）,热带念珠菌4株（5.26%）,克柔念珠菌2株（2.63%）,近平滑念珠菌1株（1.32%）。非白色念珠菌中以光滑念珠菌为主,在RVVC组中的比例明显高于VVC组,有显著性差异（P〈0.05）。药敏试验显示,RVVC的念珠菌株中仅8株对7种药物全部敏感,23株敏感药物〈5种,其中1株仅对制霉菌素敏感;对药物的敏感率为制霉菌素（97.37%）〉克霉唑（60.52%）〉酮康唑（51.32%）〉伊曲康唑（36.84%）〉咪康唑（35.53%）〉氟康唑（23.68%）〉特比奈芬（10.53%）。结论：白色念珠菌仍是RVVC的主要致病菌,非白色念珠菌比例在RVVC组中显著高于VVC组,其中以光滑念珠菌为主。念珠菌对制霉菌素敏感率最高,对唑类药物耐药性有增加趋势。     

阴道分泌物酵母样真菌分离培养及其药敏的临床价值

目的了解阴道分泌物酵母样真菌的分离培养及其药敏临床意义。方法从阴道分泌物中分离到的196株酵母样真菌,采用API-20Aux和ATBFUN GUS进行鉴定和药敏试验,所得结果进行统计分析。结果196株酵母样真菌中,分离率最多的前3种是白色念珠菌（80．5％）、葡萄牙念珠菌（9．5％）和光滑念珠菌（5．6％）,酵母样真菌对两性霉素B和制霉菌素敏感率最高,达94．5％和92．6％,5-氟胞嘧啶、咪康唑、益康唑和酮康唑只有43．6％～58．1％。结论真菌性阴道炎主要以白色念珠菌感染为主,感染的酵母菌除了对多烯类药物敏感率高外,对其他抗真菌药有不同程度的下降,应引起关注。     

Identification and susceptibility against fluconazole and albaconazole of 100 yeasts' strains isolated from vaginal discharge

Vulvovaginal candidiasis is a condition that affects a great number of fertile women. It is considered the second cause of genital infection after vaginosis due to GAM complex. Candida albicans is the most frequent isolated species from vaginal discharge. However, sometimes more than one yeast species could be found in the same clinical sample that are more resistant to antifungal drugs. Nowadays, it is necessary to identify properly up to species level the isolated microorganism and to determine the antifungal susceptibility profile. One hundred strains obtained from vaginal discharge of 94 patients suffering acute vulvovaginal candidiasis were studied. The identification of the isolates showed: C. albicans 86%, Candida glabrata 6%, Candida inconspicua 3%, Candida krusei 2% and Candida intermedia, Candida holmii and Trichosporon asahii one case each. Minimal inhibitory concentrations (MIC) of all the yeasts against fluconazole and albaconazole were performed. C. glabrata, C. krusei and C. inconspicua were the most resistant against fluconazole, on the other hand albicans was susceptible to this drug. All the isolates presented MIC against albaconazole much lower than fluconazole.     

A colorimetric assay for the assessment of cytotoxicity of yeasts

A colorimetric assay for the quantitation of microbial cytotoxicity has been developed using cells from a monocyte-like human cell line (U937), epithelial cells (Hela), and fibroblast-like cells (Vero) as targets. The fraction of surviving cells was determined by their content of the dye neutral red which is retained only by live cells and can be quantitated photometrically after controlled lysis. The neutral red retention assay was at least as sensitive as the 51Cr-release assay; it was considerably less laborious, faster, and avoided handling of radioactivity. Among the different Candida species tested, the highest cytotoxicity was associated with C. albicans and C. tropicalis; a lower degree of cytotoxicity was exhibited by C. glabrata, C. guilliermondii, C. krusei, C. parapsilosis, and C. pseudotropicalis. Among the strains of a given fungal species cytotoxicity varied by up to 40%.     

in Vitro Activity of Antimycotic Agents Determined by e-Test Method Against Vaginal Shape Candida Species

Vaginal candidiasis continues to be a common cause of vaginal discharge, pruritus and other local complaints in women worldwide. Although numerous antimycotic agents are available for the treatment of yeast vaginitis there is little comparative data on the in vitro activity of these drugs. The objectives of this study were to isolate and identify the Candida species in the vagina and anus of patients treated in a gynaecology clinic, as well as determine the susceptibility to azolic compounds measured by the E-test method. Vaginal and rectal swabs were collected from 80 adult non-pregnant patients, seen at a gynaecological clinic, aged 18–59 years, with sexual activity, with and without vaginitis. The swabs were processed by methods routinely used for the detection of pathogenic yeasts. The susceptibility of the isolates to fluconazole, ketoconazole and itraconazole, was measured by the agar diffusion method (E-test), using RPMI 1640 medium with 2% glucose and phosphate buffer. Candida species (33) strains were isolated from 17 patients at similar proportions from both anatomical sites, and 12 patients harboured 24 strains of C. albicans in the vaginal and rectal tracts. Twenty one percent of the strains of C. albicans were resistant to ketoconazole, 54% were resistant to itraconazole and 0% were resistant to fluconazole. The sensitivity of strains isolated from the two sites were similar, indicating that these are strains of the same phenotype. This revised version was published online in August 2006 with corrections to the Cover Date.     

ATB FUNGUS2法在念珠菌属和新生隐球菌抗真菌药敏试验中应用的评价

目的评价ATBFUNGUS2半固体培养基法在测定念珠菌属和新生隐球菌对4种常用抗真菌药物敏感性中的应用价值。方法利用CLSIM27．A2微量液基稀释法和ATBFUNGUS2法同时测定131株念珠菌和20株新生隐球菌对两性霉素B（AmB）、氟康唑（FLC）、氟胞嘧啶（5-Fc）和伊曲康唑（ITC）的敏感性。结果①两种方法对于AmB、5-FC、FLC和ITC的一致性分别为98％、89．4％、78．8％和78．1％;②所有受试菌株中两种方法的一致性为80％,但ATBFUNGUS2法将2／5株M27-A2法检查为FLC耐药的白念珠菌判断为敏感或剂量依赖,将8／10株M27-A2法检查为FLC剂量依赖的白念珠菌判断为敏感或耐药。③ATBFUNGUS2法中AmB的MIC值判读范围偏高,以致于实际工作中不能读出准确的值。结论ATBFUNGUS2半固体培养基法在测定念珠菌属和新生隐球菌对4种常用抗真菌药物的敏感性时不失为简单、快速而且重复性好的方法。     

Rapid identification of medically important yeasts by electrophoretic protein patterns

A simple electrophoretic method for yeast identification was evaluated. Whole cells were extracted by SDS and the protein profiles obtained in SDS-PAGE after Coomassie blue staining were compared for 52 strains from 9 species of yeast or yeast-like fungi commonly isolated from man (Candida albicans, C. glabrata, C. guilliermondii, C. krusei, C. parapsilosis, C. pseudotropicalis, C. tropicalis, Geotrichum candidum, Saccharomyces cerevisiae). The corresponding patterns showed 30 to 45 polypeptides in the range 95-20 kDa and were clearly different for the 9 species. No differences could be detected between strains from the same species. The characteristic patterns were obtained within 24 h allowing rapid identification of the most commonly encountered clinical yeast isolates.     

Candida biotypes isolated from clinical specimens in Malaysia

The distribution of Candida species was examined using 1114 yeasts isolated from various clinical specimens. The isolates were identified by germ tube test, hyphal/pseudohyphae and chlamydoconidia production and carbohydrate assimilation test using ten carbohydrates (glucose, sucrose, trehalose, cellobiose, arabinose, galactose, mannitol, raffinose, lactose and maltose). Among the 1114 isolates studied, 9 species of Candida were identified and the relative frequency of isolation was C. albicans (44.2%), C. parapsilosis (26.0%), C. tropicalis (17.7%), C. glabrata (9.6%), C. krusei (1.2%), C. rugosa (0.6%), C. guilliermondii (0.2%), C. lusitaniae (0.08%) and C. kefyr (0.08%). Non- C. albicans was the most common Candida species isolated from blood, respiratory system, urine and skin. The isolate from vaginal swabs was predominantly C. albicans. 82.2% of C. glabrata and 64.2% of C. krusei isolated in this study were from vaginal swabs. This revised version was published online in August 2006 with corrections to the Cover Date.     

Identification and antifungal susceptibility of Candida isolated from intensive care unit patients

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%.     

The role of Candida kefyr (C. pseudotropicalis) in reproductive tract infections]

Frequency of appearance of C. kefyr strains, their diagnosis, susceptibility to drugs and importance in pathogenesis of reproductive tract mycoses were investigated. The investigated material consisted of 2717 strains, and there were 67 strains of C. kefyr isolated from vaginal mucous membrane. C. kefyr strains constituted 2.5% of total number of 2717 isolated strains and 8.2% within fungi other than C. albicans species. Vaginal mycosis caused by C. kefyr occurs in presence of Lactobacillus sp. and correct values of pH in vaginal contents, however changing leukocyte number was observed. Clinical symptoms and complaints regarding reproductive tract were present in 47.8% of patients with C. kefyr infections. Investigating susceptibility to antimycotic drugs, regardless of appointed method, low susceptibility of C. kefyr strains to amphotericin B was noted (79% resistant strains). Strain of C. kefyr were sensitive to all remaining drugs.     

Minimal inhibitory concentrations of Candida species to five antifungals by using the reference dilution micromethod and the E-test.

It is accepted that the frequency of candidosis has increased during the last decade, specially in hospitalized patients. The more frequent use of azole antifungals and the recognition of isolates of Candida sp resistant to these and other drugs such as 5-fluorocytosine constitute a great need for a reproducible and useful C. albicans in vitro susceptibility testing method for monitoring antifungal therapy in clinical mycological laboratories. The E-test is a novel agar diffussion technique for testing the susceptibility of yeasts against a defined continous gradient of drug and could be used by most clinical laboratories. In this study the E-test and the NCCLS reference microbroth method (M27-P guidelines) were used to determine the MICs of amphotericin B, 5-flucytosine, itraconazole, fluconazole and ketoconazole for 50 clinical isolates of Candida albicans, Torulopsis glabrata, C. tropicalis and Hansenula anomala and five reference ATCC strains. The main purpose of the study was to compare the results obtained by the two methods. In general good agreement (+/- 1 dilution) was otained between both methods, despite differences observed for some species-antifungal combinations in which the MICs were lower by the E-test than by the microbroth method. MICs for C. albicans and T. glabrata to amphotericin B were < 0.50 microg/mL. Two isolates of C. albicans and two others of H. anomala, showed MIC < 8 microg/mL for 5- flucytosine. All isolates of T. glabrata and 40% of C. albicans showed MICs > 16 microg/mL for fluconazole. The results of this study indicate that E-test is an alternative for susceptibility testing to the NCCLS reference method. Because its simplicity it seems to be an easier test for routine clinical laboratories.     

Multicenter survey of in vitro antifungal resistance in yeasts of medical importance isolated from Spanish patients

Twelve Spanish laboratories collected 325 yeast clinical isolates during a 30 day's period, among them 224 Candida albicans, 30 Candida glabrata, and 27 Candida parapsilosis. In vitro antifungal susceptibility to amphotericin B, ketoconazole, fluconazole and itraconazole was determined by an agar diffusion test (Neo-Sensitabs, Rosco, Denmark). All the isolates tested were susceptible in vitroto amphotericin B and nearly all (97.2%) to itraconazole. In vitrosusceptibility to fluconazole and ketoconazole was high (90.2% and 91.4% of isolates, respectively) but showed variations depending on the species tested. Resistance to fluconazole and ketoconazole was low in C. albicans (4% and 3%, respectively), but 30% of Candida guilliermondii and 36% of C. glabrata isolates were resistant to fluconazole. Ketoconazole resistance was observed in 40% of C. glabrata, and 17% of Candida tropicalis. Resistance to antifungal drugs is very low in Spain and it is related to non-C. albicans isolates.     

Effects of Antifungal Agents Alone and in Combination Against Candida glabrata Strains Susceptible or Resistant to Fluconazole

The rise of Candida spp. resistant to classic triazole antifungal agents has led to a search for new therapeutic options. Here, we evaluated combinations of antifungals in a checkerboard assay against two groups of Candida glabrata strains: one containing fluconazole-susceptible clinical isolates (FS) and another containing fluconazole-resistant laboratory derivative (FR). The most synergistic combination observed was amphotericin B + flucytosine (synergistic for 61.77 % of FS strains and 76.47 % of FR strains). The most antagonistic combination observed was ketoconazole + flucytosine (FS 61.77 % and FR 55.88 %). Surprisingly, most combinations evidenced indifferent interactions, and the best synergism appeared when amphotericin B and flucytosine were combined against both groups of isolates.     

Synergistic anticandidal activity of xanthorrhizol in combination with ketoconazole or amphotericin B

Candida species are responsible for the fourth most common nosocominal bloodstream infection. Xanthorrhizol, a sesquiterpene compound isolated from Curcuma xanthorrhiza Roxb. has been reported to have anticandidal activity. The aim of this study is to investigate the synergistic anticandidal effect of xanthorrhizol in combination with ketoconazole or amphotericin B against Candida albicans, Candida glabrata, Candida guilliermondii, Candida krusei, Candida parapsilosis , and Candida tropicalis . Mostly, xanthorrhizol in combination with ketoconazole or amphotericin B exhibited the synergistic anticandidal effects against all species of Candida tested. In combination with xanthorrhizol, the concentration of ketoconazole or amphotericin B for inhibiting the growth of the tested Candida species could be reduced by ≥50%. Time–kill curves showed that 1/2 minimum inhibitory concentration (MIC) dose of xanthorrhizol, amphotericin B, or ketoconazole alone against each of the six Candida species did not inhibit the growth of all Candida species tested. However, 1/2 MIC dose of xanthorrhizol in combination with 1/2 MIC dose of ketoconazole or 1/2 MIC dose of amphotericin B exhibited growth inhibition of all Candida species tested and reduced viable cells by several logs within 4 h. These results support the potential use of xanthorrhizol as an anticandidal agent, and it can be used complementarily with other conventional antifungal agents.