Antifungal drug susceptibility of Cryptococcus neoformans from clinical sources in Nairobi, Kenya

The serotypes and mating types of 80 clinical isolates of Cryptococcus neoformans from Kenya were studied and subjected to broth microdilution susceptibility testing to amphotericin B (AMP), flucytosin, fluconazole (FLC), itraconazole (ITC) and miconazole (MCZ). The isolates included C. neoformans var. grubii- 75 of 80 (serotype A; 93.7%), C. neoformans var. neoformans- three of 80 (3.8%) and C. neoformans var. gattii- two (serotype B; 2.5%). Mating experiment confirmed all the isolates to be alpha-mating type. Seventy-eight (97.5%) of the isolates had minimum inhibitory concentration (MIC) of < or =0.5 microg ml(-1) to AMP and at 1 microg ml(-1), 100% of the isolates were inhibited. Flucytosin resistance was observed in 21% with MIC in which 90% of the isolates were inhibited (MIC90) of 64 microg ml(-1). Only 23.8% of the strains were susceptible to FLC with 65% susceptible dose-dependent (SDD) and 11.2% resistant. Itraconazole susceptibility was 61.3% while the rest were either SDD or resistant. The MIC90 for ITC and MCZ were 0.5 and 2 microg ml(-1) respectively. The study reports the serotypes, mating types and highlights the existence of azoles resistance in C. neoformans in Nairobi which calls for antifungal drug resistance surveillance as prophylactic use of FLC increases because of human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS) epidemic in sub-Saharan Africa.     

Evaluation of antifungal combination against Cryptococcus spp.

The second cause of death among systemic mycoses, cryptococcosis treatment represents a challenge since that 5‐flucytosine is not currently available in Brazil. Looking for alternatives, this study evaluated antifungal agents, alone and combined, correlating susceptibility to genotypes. Eighty Cryptococcus clinical isolates were genotyped by URA5 gene restriction fragment length polymorphism. Antifungal susceptibility was assessed following CLSI‐M27A3 for amphotericin (AMB), 5‐flucytosine (5FC), fluconazole (FCZ), voriconazole (VRZ), itraconazole (ITZ) and terbinafine (TRB). Drug interaction chequerboard assay evaluated: AMB + 5FC, AMB + FCZ, AMB + TRB and FCZ + TRB. Molecular typing divided isolates into 14 C. deuterogattii (VGII) and C. neoformans isolates were found to belong to genotype VNI (n = 62) and VNII (n = 4). C. neoformans VNII was significantly less susceptible than VNI (P = 0.0407) to AMB; C. deuterogattii was significantly less susceptible than VNI and VNII to VRZ (P < 0.0001). C. deuterogattii was less susceptible than C. neoformans VNI for FCZ (P = 0.0170), ITZ (P < 0.0001) and TRB (P = 0.0090). The combination FCZ + TRB showed 95.16% of synergistic effect against C. neoformans genotype VNI isolates and all combinations showed 100% of synergism against genotype VNII isolates, suggesting the relevance of cryptococcal genotyping as it is widely known that the various genotypes (now species) have significant impact in antifungal susceptibilities and clinical outcome. In difficult‐to‐treat cryptococcosis, terbinafine and different antifungal combinations might be alternatives to 5FC.     

Antifungal susceptibility profile in vitro of Sporothrix schenckii in two growth phases and by two methods: microdilution and E‐test

The susceptibility profile of 91 Sporothrix schenckii isolates in both growth phases was determined by microdilution test (Antifungal Susceptibility Testing of the European Committee for Antimicrobial Susceptibility Testing; AFST‐EUCAST). Amphotericin B (AMB), itraconazole (ITC), posaconazole, ravuconazole and terbinafine were found active in vitro against both phases but minimum inhibitory concentrations values for mycelial phase were significantly higher. Fluconazole (FLC) and voriconazole (VRC) were inactive in vitro against both phases. The E‐test technique was also performed with 41 representative isolates for AMB, FLC, ITC and VRC. Average agreement rates between yeast phase microdilution results and E‐test results were high for AMB (77.5%) and FLC (87.8%), but low for ITC and VRC with rates of 56.4% and 54.5%, respectively. AFST‐EUCAST is not the most recommended test to perform drug susceptibility testing of S. schenckii in clinical laboratories, and E‐test could be an alternative methodology for this purpose, mainly when the activity in vitro of antifungal agents of AMB and FLC are evaluated.     

Susceptibility of sequential Fonsecaea pedrosoi isolates from chromoblastomycosis patients to antifungal agents

Fourteen Fonsecaea pedrosoi isolates from six chromoblastomycosis patients were submitted to susceptibility testing. Some patients were undergoing treatment with itraconazole (ITZ) and/or cryosurgery with liquid nitrogen. The antifungal agents amphotericin B (AMB), ITZ, fluconazole (FCZ), ketoconazole (KCZ), 5-fluorocytosine (5-FC), and terbinafine (TBF) were tested. AMB and FCZ showed less activity for all isolates. The most active agents were KCZ and TBF. Sequentially isolates from four patients presented ITZ minimal inhibitory concentration (MIC) higher than the previous ones; for two of these patients, response to therapy with this agent was not observed. These results suggest development of microbiologic resistance to ITZ in four instances, two of them coinciding with lack of clinical response to this drug.     

Molecular typing and antifungal susceptibility of clinical and environmental Cryptococcus neoformans species complex isolates in Goiania, Brazil

A total of 124 Cryptococcus isolates, including 84 clinical strains obtained from cerebrospinal fluid from AIDS patients and 40 environmental isolates from pigeon excreta and from Eucalyptus trees, were studied. The varieties, serotypes, phospholipase activity and molecular profile of these isolates were determined. Cryptococcus neoformans var. grubii serotype A was identified in 120 isolates and Cryptococcus gattii serotype B in four isolates. The clinical isolates showed higher phospholipase activity than environmental isolates. Similar patterns of in vitro susceptibility to amphotericin B, fluconazole, itraconazole and voriconazole and no resistance were found for all isolates. Molecular type VNI ( C. neoformans var. grubii ) was recovered in 80 clinical and 40 environmental isolates while the type VGIII ( C. gattii ) was found in four clinical isolates. This study demonstrated for the first time the molecular types of clinical and environmental Cryptococcus isolates in the midwest Brazil region.     

肿瘤化疗患者假丝酵母菌感染的研究

目的:研究住院肿瘤化疗患者中假丝酵母菌感染的分布及耐药性,为临床合理应用抗真菌药物提供依据。方法:用法国生物梅里埃公司提供的ID 32C做假丝酵母菌的鉴定,按CLSI要求,用ATBTM Fungus真菌药敏试验板条进行药敏试验。结果:假丝酵母菌属感染的主要真菌有5种,其中以白色假丝酵母菌最多,占55.3%;其他依次为热带假丝酵母菌占13.0%,光滑假丝酵母菌占12.6%,近平滑假丝酵母菌占8.9%,克柔假丝酵母菌占4.8%;假丝酵母菌耐药吡咯类较普遍,5-氟胞嘧啶次之(17.3%),对两性毒素B耐药(2.5%)的较少。结论:假丝酵母菌属感染主要由白色假丝酵母菌引起;假丝酵母菌对目前抗真菌药呈普遍耐药性,应联合足量用药。     

In vitro susceptibilities of Malaysian clinical isolates of Cryptococcus neoformans var. grubii and Cryptococcus gattii to five antifungal drugs

The in vitro susceptibilities of Malaysian clinical isolates of Cryptococcus neoformans var. grubii and C . gattii to five antifungal drugs (amphotericin B, flucytosine, fluconazole, itraconazole and ketoconazole) were determined using the Etest method. None of the Malaysian isolates was resistant to amphotericin B and ketoconazole. Isolates resistant to flucytosine, fluconazole and itraconazole were observed in this study. Minimum inhibition concentrations (MICs) of > or = 32 microg ml(-1) against flucytosine, > or = 64 microg ml(-1) against fluconazole and > or = 1 microg ml(-1) against itraconazole were noted in four (8.3%), two (4.2%) and one (2.1%) isolates respectively. There was no significant difference in the MICs for both Cryptococcus species (P > 0.05), indicating that C. gattii was as susceptible as var. grubii to all the antifungal drugs tested. No significant difference in the MICs for both Cryptococcus species collected from 1980 to 1990 and 2002 to 2004 were observed (P > 0.05).     

Susceptibility of yeast isolates from defined German patient groups to 5-fluorocytosine.

Susceptibility and development of resistance to 5-fluorocytosine (5-FC) in Candida strains isolated from defined German groups of probands was investigated. 5-FC susceptibility was determined in a microdilution assay in yeast nitrogen base after 24 h of incubation at 37 degrees C. The range of 5-FC concentration investigated was between 0.015 and 16 micrograms ml-1. Isolates with a minimum inhibitory concentration (MIC) of > 16 micrograms ml-1 were regarded as 5-FC resistant. In total 335 Candida isolates were investigated, and 20 of them (6.0%) were found to be resistant. The Candida isolates were rather different with respect to their origin: out of 57 vaginal isolates from non-risk patients from Southern Germany 3.5% were 5-FC-resistant strains. One hundred and fifty-nine isolates from the urine of long-term intensive care patients from the whole of Germany showed 5-FC resistance (6.3%). Out of 74 isolates of different localization from intensive care patients of the University Clinics of Freiburg, 10.8% showed 5-FC resistance. Among 45 isolates from the oral cavity of HIV-positive patients from the Frankfurt region, no 5-FC-resistant strain was found. The epidemiology of 5-FC resistance is based mainly on the percentage of non-albicans isolates of the proband groups (C. tropicalis, C. krusei and others), and is less based on the frequency of Candida albicans serotype B isolates. In sequential observations with individual intensive care patients, no increase of 5-FC resistance in their Candida isolates could be observed with longer periods of hospitalization.     

Sertaconazole: in-vitro antifungal activity against vaginal and other superficial yeast isolates.

The in vitro susceptibilities of 183 clinical yeast isolates to sertaconazole (STZ) were compared to their susceptibilities to clotrimazole (CTZ), econazole (ECZ), ketoconazole (KTZ), miconazole (MNZ), fluconazole (FLZ), itraconazole (ITZ), tioconazole (TCZ), amphotericin B (AMB) and flucytosine (5FC) by using a commercial agar diffusion method. Strains were isolated from vaginal and other superficial clinical samples (18 species of Candida and five strains belonging to other yeast genera). Only one strain (0.5%) was resistant to STZ out of 87.4% of susceptible strains (n=160). The percentage of susceptible strains was higher than those obtained with the other agents evaluated and the percentage of resistant strains was lower than for most of the other antifungals. The pattern of susceptibility of C. albicans to STZ, TCZ, ITZ and CLZ was similar and superior to the pattern of susceptibility of this species to MNZ, ECZ, FLZ, 5FC and KTZ. C. dubliniensis was more susceptible to STZ, MNZ, MNZ, FLZ, ITZ, CLZ than to TCZ, ECZ, 5FC, AMB or KTZ. Ten susceptible strains to STZ were resistant to FLZ and one strain was resistant to ITZ. The overall antifungal activity of STZ in vitro against a wide range of clinically important yeasts from vaginal and cutaneous samples indicates the therapeutic potential of this agent for the treatment of infections caused by these fungi. However, the activity of STZ and the clinical value of in vitro data need to be verified in human clinical trials.     

In vitro efficacy of amphotericin B, 5‐fluorocytosine,fluconazole, voriconazole and posaconazole against Candida dubliniensis isolates using time‐kill methodology

Candida dubliniensis is a recently described yeast that causes infections in mucosal surfaces as well as sterile body sites. Candida dubliniensis develops resistance to fluconazole (FLC) more rapidly than the closely related species C. albicans. The killing activity of amphotericin B (AMB), 5‐fluorocytosine (5FC), FLC, voriconazole (VRC) and posaconazole (POS) was determined against six C. dubliniensis clinical isolates, identified using molecular biological methods and C. dubliniensis CD36 reference strain. Minimum inhibitory concentrations (MICs) were determined using the Clinical and Laboratory Standards Institute standard procedure. Time‐kill assays were performed using RPMI‐1640 as test media over a 48‐h period. AMB proved to be fungicidal at ≥0.5 μg ml^?1 against all clinical isolates after 48 h. 5FC was only fungicidal at 32–64× MIC (4–8 μg ml^?1) against all C. dubliniensis isolates. FLC, VRC and POS were fungistatic; decrease in colony number was observed only at the highest concentrations tested (8, 4 and 4 μg ml^?1, respectively). Triazoles invariably showed fungistatic effect at concentrations attainable in the serum. In clinical situations when a fungicidal antifungal is desirable, AMB may be used.     

Current scenario of cryptococcosis and antifungal susceptibility pattern in India: a cause for reappraisal

This study analysed the spectrum, antifungal susceptibility pattern, clinical course and molecular epidemiology of cryptococcosis. Four hundred and thirty-nine samples obtained from 378 meningitis patients were processed by standard procedures. Minimum inhibitory concentration (MIC) of fluconazole and amphotericin B for the isolates was tested by broth micro dilution and by E-strip method. Molecular analysis by random amplified polymorphic DNA-PCR of eight isolates was performed using M13 primer. Cryptococcosis was diagnosed in 35 patients [HIV-1 seropositive (19) and apparently immunocompetent (16)]. Cryptococcus neoformans var. neoformans (serotype A and D) was the predominant isolate on phenotypic identification. Three C. neoformans var. gattii were isolated from HIV-1 seropositive (2) and apparently immunocompetent (1) patients. MIC 90 for amphotericin B and fluconazole were 1 and 8 μg ml⁻¹ respectively. On RAPD-PCR, less diversity was seen among Indian isolates. AIDS remains the single most important risk factor for cryptococcosis. Rising MIC of the available induction and maintenance drugs is of grave concern. The DNA typing technique showed less diversity among Indian strains. Routine surveillance and application of molecular typing methods are crucial to know the baseline and existing pattern of cryptococcosis.     

Serotype identification of Cryptococcus neoformans by multiplex PCR

The pathogenic yeast Cryptococcus neoformans is traditionally classified into three varieties with five serotypes: var. grubii (serotype A), var. neoformans (serotype D), var. gattii (serotypes B and C) and serotype AD (hybrid of serotypes A and D). A commercial kit, Crypto Check (Iatron Laboratories, Tokyo, Japan), has been used worldwide for serotyping isolated strains. However, its production was discontinued in 2004, and hence the present study aimed to develop a simple polymerase chain reaction (PCR) method for serotyping C. neoformans strains. Subjecting genomic DNA of 59 strains of the five serotypes to multiplex PCR amplification using a set of four primers designed for the laccase gene (LAC1) differentiated serotypes A, D, B and C, but could not separate serotype AD from serotype D. However, a primer pair designed for the capsule gene (CAP64) allowed serotypes D and AD to be differentiated. When PCR amplification was performed in the simultaneous presence of the above six primers, the five serotypes produced two to five DNA fragments that could be used to distinguish them. This multiplex PCR method is useful for serotyping C. neoformans isolates, and represents an effective replacement for the Crypto Check kit.     

肿瘤化疗患者假丝酵母菌感染的研究

目的：研究住院肿瘤化疗患者中假丝酵母菌感染的分布及耐药性,为临床合理应用抗真菌药物提供依据。方法：用法国生物梅里埃公司提供的ID 32C做假丝酵母菌的鉴定,按CLSI要求,用ATBTM Fungus真菌药敏试验板条进行药敏试验。结果：假丝酵母菌属感染的主要真菌有5种,其中以白色假丝酵母菌最多,占55.3%;其他依次为热带假丝酵母菌占13.0%,光滑假丝酵母菌占12.6%,近平滑假丝酵母菌占8.9%,克柔假丝酵母菌占4.8%;假丝酵母菌耐药吡咯类较普遍,5-氟胞嘧啶次之（17.3%）,对两性毒素B耐药（2.5%）的较少。结论：假丝酵母菌属感染主要由白色假丝酵母菌引起;假丝酵母菌对目前抗真菌药呈普遍耐药性,应联合足量用药。     

Follow-up of epidemiological data of cryptococcosis in Austria, Germany and Switzerland with special focus on the characterization of clinical isolates

The present survey in Austria, Germany and Switzerland continued the survey of cryptococcosis set up by the European Confederation of Medical Mycology (ECMM) in 1997. From 2000 to 2003 77 cases have been reported. An HIV infection is still the most important risk factor (68%). Young HIV+ women from ASIA contributed to the increase of cryptococcosis in females. A total of 129 clinical isolates of both surveys were genotyped by PCR fingerprinting to study the prevalence of different genotypes. The prevalence of Cryptococcus neoformans var. grubii (serotype A) with the genotypes VNA1 and VNA2 was higher in Germany and Austria (74.5%) than in Switzerland (52%), while in Switzerland the Cr. neoformans hybrids AD (26%) and Cr. neoformans var. neoformans (serotype D) (22%) were more prevalent compared with Germany and Austria (8 and 17.5% respectively). Cryptococcus gattii isolates were studied by FT-IR spectroscopy. DNA in the ITS region was sequenced to get further information about Cr. neoformans serotype AD strains and about the geographical origin of the Cr. gattii isolates. The ITS sequence of the serotype AD isolates of the genotypes VNAD1, VNAD2 and VNAD4 is usually identical to serotype A or serotype D respectively. In the three isolates of the genotype VNAD3 a genotype-specific sequence pattern was detected. Two autochthonous infections due to Cr. gattii could indicate that the genotype VGIV with the ITS type 'Asia 2' might be endemic in Europe.     

In vitro susceptibility of Cryptococcus neoformans serotypes to GM 237354 derivative of the sordarin class

In vitro susceptibility to the sordarin derivative GM 237354 and amphotericin B were tested in a total of 190 Cryptococcus neoformans clinical isolates from different geographical areas of Spain and South American countries. Minimal inhibitory concentrations (MICs) were obtained using the NCCLS reference microbroth dilution method and analysed according the serotypes of Cr. neoformans. The MICs for amphotericin B were lower than 1.0 microg ml(-1) (MIC90% 0.5 microg ml(-1) , MIC50% 0.125 microg ml(-1)) but five isolates showed MICs of 2.0 microg ml(-1) to GM 237354 (MIC90% 1.0 microg ml(-1), MIC50% 0.5 microg ml(-1)). Cryptococcus neoformans var. gattii serotype B, was significantly less susceptible than A and AD serotypes (P = 0.047 and P = 0.022, respectively).     

Sertaconazole: In-Vitro Antifungal Activity Against Vaginal and Other Superficial Yeast Isolates

Abstract

The in vitro susceptibilities of 183 clinical yeast isolates to sertaconazole (STZ) were compared to their susceptibilities to clotrimazole (CTZ), econazole (ECZ), ketoconazole (KTZ), miconazole (MNZ), fluconazole (FLZ), itraconazole (ITZ), tioconazole (TCZ), amphotericin B (AMB) and flucytosine (5FC) by using a commercial agar diffusion method. Strains were isolated from vaginal and other superficial clinical samples (18 species of Candida and five strains belonging to other yeast genera). Only one strain (0.5%) was resistant to STZ out of 87.4% of susceptible strains (n=160). The percentage of susceptible strains was higher than those obtained with the other agents evaluated and the percentage of resistant strains was lower than for most of the other antifun-gals. The pattern of susceptibility of C. albicans to STZ, TCZ, ITZ and CLZ was similar and superior to the pattern of susceptibility of this species to MNZ, ECZ, FLZ, 5FC and KTZ. C. dubliniensis was more susceptible to STZ, MNZ, MNZ, FLZ, ITZ, CLZ than to TCZ, ECZ, 5FC, AMB or KTZ. Ten susceptible strains to STZ were resistant to FLZ and one strain was resistant to ITZ. The overall antifungal activity of STZ in vitro against a wide range of clinically important yeasts from vaginal and cutaneous samples indicates the therapeutic potential of this agent for the treatment of infections caused by these fungi. However, the activity of STZ and the clinical value of in vitro data need to be verified in human clinical trials.     

Incidence of cryptococcosis in and around Chandigarh, India, during the period 1982-91

Summary. The incidence of cryptococcosis was evaluated in and around Chandigarh, India over a period of 10 years (1982-91). Different species of Cryptococcus were isolated from 38 patients. Cryptococcus neoformans was the predominant isolate (26 = 68%) followed by Cr. albidus (2), Cr. laurentii (1) and not precisely identified Cryptococcus species (9 = 24%). Serotyping of the 18 isolates of Cr. neoformans revealed that 13 (72%) were serotype A, two (11%) were serotype AD and three (17%) were serotype B ( Cr. neoformans var. gattii). Cryptococcus species were found to produce infection in 24 patients and were possibly transient colonizers in another 14 patients. In addition, in 11 patients no Cryptococcus species was isolated from any site but the latex agglutination test for antigen (Crypto-LA) was positive in serum and/or cerebrospinal fluid. Response to various antifungal drugs was also studied in these patients.
Zusammenfassung. Über den Zehnjahreszeitraum 1982–1991 wurde in der Region Chandigarh, Indien, die Cryptococcose-Häufigkeit erhoben. Bei 38 Patienten wurde Cryptococcus isoliert. Vorherrschend war mit 26 Isolaten (68%) Cryptococcus neoformans , gefolgt von Cr. albidus (2), Cr. laurentii (1) und nicht näher definierten Cryptococcus -Arten (9 Isolate = 24%). Bei der Serotypisierung von Cr. neoformans erwiesen sich 13 (72%) als zum Serotyp A gehörig, zwei (11%) gehörten zum Serotyp AD und drei (17%) zum Serotyp B ( Cr. neoformans var. gattii). Cryptococcus -Arten führten bei 24 Patienten zu Infektionen und bei weiteren 14 Patienten möglicherweise zu einer Kolonisation. Bei 11 Patienten konnte Cryptococcus nicht nachgewiesen werden, aber der Cryptococcus -Antigennachweis (Crypto-LA Latexagglutination) war im Serum und/oder im Liquor positiv. Das Ansprechen auf unterschiedliche Antimykotika wurde gleichfalls in die Studie einbezogen.     

Identification of Candida species isolated from patients in intensive care unit and in vitro susceptibility to fluconazole for a 3-year period

Species level identification of Candida and antifungal susceptibility testing is not generally performed in routine laboratory practice. There is limited information about the distribution of Candida species and antifungal susceptibility in Turkey. In this study, we aimed at identifying Candida isolates to species level from various samples obtained from patients treated in an intensive care unit between 2002 and 2005 and to evaluate fluconazole susceptibilities of the isolates. A total of 320 Candida isolates obtained from 270 patients were identified by conventional methods and using API (Candida and/or 20C AUX) system. Antifungal susceptibility testing was performed by broth microdilution method. Candida albicans was isolated with the highest frequency (65.6%) followed by C. parapsilosis (11.3%), C. glabrata (8.8%) and C. tropicalis (7.8%). Of all the isolates, 92.9% revealed susceptibility to fluconazole. Susceptibility to fluconazole was highest for C. albicans followed by C. parapsilosis and C. glabrata. The MIC(90) values for C. albicans, C. parapsilosis, C. glabrata and C. tropicalis were 1, 2, 8 and 4 mug ml(-1) respectively. Fluconazole remains effective against both C. albicans and the majority of non-albicans Candida species. In this study, we determine the distribution of Candida species and evaluate the susceptibilities of the isolates, particularly for the azoles.     

Effects of several antifungal drug combinations against clinical and environmental isolates of Cryptococcus neoformans from China

The in vitro interactions of caspofungin (CSP) with terbinafine (TRB) and ravuconazole (RVC) with 5-fluorocytosine (5-FC) were tested against 82 clinical and environmental isolates of Cryptococcus neoformans from China. The interaction of CSP with TRB proved synergistic against those isolates with a CSP MIC < or =2 microg ml-1 (5% of the isolates), additive against 42% of the isolates and indifferent against 53%. The effects of RVC with 5-FC were synergistic, additive or indifferent against 8%, 26% and 67% of the isolates, respectively. No antagonistic effects were found among any of the drugs. The combinations of CSP with TRB and RVC with 5-FC may display beneficial effects in a strain-dependent manner, while in no case showed antagonistic effects. These data might be of use to design safer and more efficient treatments for patients with cryptococcosis and warrant further evaluation.     

In vitro antifungal activity of Indian liposomal amphotericin B against clinical isolates of emerging species of yeast and moulds,and its comparison with amphotericin B deoxycholate,voriconazole, itraconazole and fluconazole

Fungisome^TM is a liposomal preparation of amphotericin B (AMB), already marketed in India. However, its antifungal activity has not been evaluated against a wide range of fungal pathogens. The study was planned to elucidate the in vitro antifungal activity of Fungisome^TM against wide range of fungi and compare it with AMB deoxycholate (AMB‐d), voriconazole (VOR), itraconazole (ITR) and fluconazole (FLU). Minimum inhibitory concentrations (MICs) of the drugs were determined for 262 clinical fungal isolates, including yeast, dimorphic and filamentous fungi, by broth microdilution method approved by Clinical and Laboratory Standards Institute, USA (yeast, M27‐A3; filamentous fungi, M38‐A2). The MIC_90s of Fungisome^TM were 0.125, 0.5 and 0.25 mg l^?1 against yeast, filamentous and dimorphic fungi respectively. In comparison, MIC_90s of AMB‐d, FLU, ITR and VOR were 1, 1 and 1 mg l^?1 (AMB‐d), 4, 64 and 64 mg l^?1 (FLU), 1, 16 and 16 mg l^?1 (ITR) and 0.5, 4 and 16 mg l^?1 (VOR) against yeast, filamentous and dimorphic fungi respectively. The MIC of Fungisome^TM was two to 16‐fold lower than AMB‐d. These results reveal an efficient in vitro activity of Fungisome^TM.