Antifungal Activity of Diethyldithiocarbamate Antimyzetische Aktivitat von Diethylcarbamat

Sodium diethyldithiocarbamate (DTC) was evaluated for its ability to combat four different species of fungi in vitro. Using a microtiter-broth-dilution method we were able to demonstrate an antifungal activity against Candida albicans, Cryptococcus neoformans, Aspergillus fumigatus and Mucor mucedo in doses achievable by intravenous administration in man.     

Caspofungin: when and how? The microbiologist’s view

The echinocandins are antifungal agents, which act by inhibiting the synthesis of β-(1,3)-D-glucan, an integral component of fungal cell walls. Caspofungin, the first approved echinocandin, demonstrates good in vitro and in vivo activity against a range of Candida species and is an alternative therapy for Aspergillus infections. Caspofungin provides an excellent safety profile and is therefore favoured in patients with moderately severe to severe illness, recent azole exposure and in those who are at high risk of infections due to Candida glabrata or Candida krusei. In vivo/in vitro resistance to caspofungin and breakthrough infections in patients receiving this agent have been reported for Candida and Aspergillus species. The types of pathogens and the frequency causing breakthrough mycoses are not well delineated. Caspofungin resistance resulting in clinical failure has been linked to mutations in the Fksp subunit of glucan synthase complex. European Committee for Antimicrobial Susceptibility Testing and Clinical and Laboratory Standards Institute need to improve the in vitro susceptibility testing methods to detect fks hot spot mutants. Caspofungin represents a significant advance in the care of patients with serious fungal infections.     

Epidemiological and mycological data of onychomycosis in Goiania, Brazil

Onychomycosis defined as fungal infection of the nail represents more than 50% of all onychopathies. Epidemiological studies have shown that this mycosis is worldwide in occurrence, but with geographical variation in distribution. The direct microscopy and culture of the nail samples were performed to identify the causative agent. Out of 2273 patients with nail infection examined between January 2000 and December 2004 in Goiania, state of Goias, Brazil, diagnosis of onychomycosis was confirmed in 1282 cases, with dermatophytes and Candida species being the most common aetiological agents isolated. Dermatophyte onychomycosis was more common in toenails than in fingernails, while onychomycosis caused by yeast had a similar frequency in both toenails and fingernails. Among the species identified, Candida albicans was responsible for 492 cases (38.4%) of onychomycosis, Trichophyton rubrum was found in 327 cases (25.6%) and Trichophyton mentagrophytes in 258 cases (20.1%). Other fungi isolated from nail infections included Aspergillus sp., Trichosporon sp., Geotrichum sp. and Fusarium sp. In our study, yeast of the genus Candida were the dominant cause of onychomycosis in women and dermatophytes were the principal cause of this condition in men.     

Species assignment and antifungal susceptibilities of black aspergilli recovered from otomycosis cases in Iran

Black aspergilli are among the main causative agents of otomycosis worldwide. In this study, the species assignment of black aspergilli isolated from otomycosis cases in Iran was carried out using sequence analysis of part of the calmodulin gene. The results indicate that Aspergillus niger is not the only black Aspergillus species involved in otomycosis cases in Iran: Aspergillus awamori and Aspergillus tubingensis are also able to cause ear infections. Antifungal susceptibility tests were carried out against five antifungal drugs including amphotericin B, fluconazole, itraconazole, ketoconazole and terbinafine. All isolates were highly susceptible to terbinafine, while they exhibited moderate susceptibilities against amphotericin B, fluconazole and ketoconazole. Aspergillus niger and A. awamori were found to have higher minimal inhibitory concentrations for azoles than A. tubingensis, in accordance with previous findings.     

Aspergillus and Candida Species Isolated from the Sputa of Patients with Bronchopulmonary Disorders in Nigeria

Summary: Sputum specimens from 100 patients with bronchopulmonary disorders were investigated for micro-organisms of etiologic significance. Screening tests were put up for fungi. In the process, it was observed that a large number of these specimens contained species of fungi occasionally associated with bronchopulmonary diseases. Aspergillus species, mainly A. fumigatus and A. niger were isolated from 27 cases while Candida albicans grew in 20 cases. From 13 cases such fungi as Rhizopus, Mucor and other species of doubtful significance were grown. Possible implications of these findings and the role of the diagnostic laboratory in the diagnosis of fungal infections of the lung are discussed.
Zusammenfassung: Sputumproben von 100 Patienten mit bronchopulmonalen Erkrankungen wurden auf ätiologisch bedeutende Mikroorganismen untersucht. Außerdem wurden Pilzkulturen angelegt. Dabei zeigte sich, daß eine große Zahl von Pilzen gefunden wurde, die gelegentlich mit bronchopulmonalen Erkrankungen in Verbindung gebracht werden. Aspergillusarten, vor allem A. fumigatus und A. niger wurden bei 27 Patienten isoliert, während Candida albicans bei 20 Fällen wuchs. Von 13 Patienten wurden Pilze wie Rhizopus, Mucor und andere Arten gezüchtet, deren Rolle unklar ist. Die möglichen Bedeutungen dieser Befunde und die Rolle des diagnostischen Laboratoriums bei der Diagnose von Lungenmykosen werden diskutiert.     

Inhibitory effect of terbinafine on reactive oxygen species (ROS) generation by Candida albicans

Candida albicans, the most important opportunistic fungal pathogen, is able to generate remarkable amounts of reactive oxygen species (ROS). Since ROS are highly cytotoxic, this mechanism may contribute to the pathogenicity of this yeast, including its invasiveness and the inflammatory response of the host. Terbinafine, a synthetic antifungal agent of the allylamine class, inhibits ergosterol biosynthesis at the level of squalene epoxidase. Furthermore, there is evidence that terbinafine at therapeutic concentrations can be considered a free radical scavenger in vitro and could exert an anti-inflammatory activity in vivo. In this study we investigated whether terbinafine affects the generation of ROS by C. albicans. Blastoconidia of the C. albicans strain 3153A were cultured in YEPG-medium and, subsequently, incubated with different doses of terbinafine (1, 10 and 100 microg ml(-1)) for 10 and 60 min, respectively. ROS generation was measured by lucigenin-enhanced chemiluminescence. Formation of ROS was considerably dependent on cell number. Chemiluminescence signals were measured at a concentration > or = 1 x 10(6) cells ml(-1), with a maximum of 1 x 10(8) cells ml(-)1. Already after 10 min of incubation with terbinafine, a dose-dependent significant inhibition of ROS generation was found (P < 0.05), whereas after 60 min this effect was amplified. In conclusion, terbinafine reduced the ability of C. albicans to generate ROS. Besides the known effect on ergosterol biosynthesis, this mechanism may contribute to its antifungal action.     

In vitro activity of nitroxoline against clinical isolates of Candida species

The in vitro antifungal activity of the quinoline nitroxoline has been compared with those of amphotericin B, flucytosine, and two azoles, miconazole and ketoconazole, against clinical isolates of Candida spp. A total of 186 isolates of 10 species of Candida and two culture collection strains were tested by an agar-dilution technique. Nitroxoline was highly active against Candida spp. MICs for nitroxoline ranged between 0.25-2 micrograms ml-1 for 186 representative strains. With MIC90 as the measure of antifungal activity, nitroxoline appeared to be superior to the imidazoles studied. Data for individual species of Candida revealed that the activities of nitroxoline and amphotericin B were generally just as effective against C. albicans, whereas flucytosine was the most active agent against Candida spp.     

Assessing risk factors for systemic fungal infections

The incidence of invasive fungal infection has increased in recent years. Most infections are caused by Candida albicans and Aspergillus spp. but the emergence of other fungal infections is changing the spectrum of disease. Immunosuppression and breakdown of anatomical barriers such as the skin are the major risk factors for fungal infections. Health care workers encounter at-risk patients in various settings, including AIDS clinics and intensive care, transplantation and oncology units. Patients with prolonged and deep neutropenia (haematological malignancy patients) are most at risk and are therefore most likely to receive prophylactic therapy. Practical measures can be taken to avoid exposing the patient to fungi (air filtration, regular hand washing, avoiding plants and flowers) and antifungal agents can be administered to prevent systemic fungal infection. Most fungal infections have non-specific symptoms; this makes recognition of the signs and symptoms of the disease important but also makes diagnosis difficult and empirical treatment necessary. Some antifungal agents have limitations but new formulations will improve therapy and play a key role in future antifungal strategies.     

Gibt es Pathogenitätsfaktoren bei Pilzen?

Zusammenfassung. Seit geraumer Zeit werden sogenannte Pathogenitäts- oder Virulenzfaktoren bei pathogenen Bakterien intensiv studiert. Während vor allem obligat pathogene Organismen, aber auch fakultativ pathogene Bakterien derartige Faktoren ausbilden, ist ihre Präsenz und Funktion bei pathogenen Pilzen nach wie vor umstritten. In der vorliegenden Arbeit werden Daten zusammengestellt, die das Wirken von Pathogenitätsfaktoren bei Cryptococcus neoformans, Aspergillus fumigatus und Candida albicans beleuchten. Es sind unter anderem Proteasen, Haftfaktoren und Kapseln, die bei Pilzen zur Pathogenitat beitragen können. Da Pilzinfektionen in der Regel bei immunsupprimierten Personen auftreten, spielt neben der Wirkung von Pathogenitätsfaktoren der Wirtsstatus eine entscheidende Rolle im Infektionsgeschehen.
Summary. Since many years so-called pathogenicity or virulence factors of pathogenic bacteria have been studied intensively. Whereas these factors are produced especially by obligate but also facultative pathogenic bacteria, their presence and function in pathogenic fungi is still a matter of debate. In this minireview we summarize some data about possible pathogenicity factors of Cryptococcus neoformans, Aspergillus fumigatus and Candida albicans. The production of adhesins, proteases and capsules contributes to the virulence of these fungi. As fungal infections usually affect immunocompromised patients, the immune status of the host plays a decisive role in addition to the virulence factors produced by fungi.     

Detection of characteristic metabolites of Aspergillus fumigatus and Candida species using ion mobility spectrometry-metabolic profiling by volatile organic compounds

Volatile metabolites of Aspergillus fumigatus and Candida species can be detected by gas chromatography/mass spectrometry (GC/MS). A multi-capillary column - ion mobility spectrometer (MCC-IMS) was used in this study to assess volatile organic compounds (VOCs) in the headspace above A. fumigatus and the four Candida species Candida albicans, Candida parapsilosis, Candida glabrata and Candida tropicalis in an innovative approach, validated for A. fumigatus and C. albicans by GC/MS analyses. For the detection of VOCs, a special stainless steel measurement chamber for the microbial cultures was used. The gas outlet was either attached to MCC-IMS or to adsorption tubes (Tenax GR) for GC/MS measurements. Isoamyl alcohol, cyclohexanone, 3-octanone and phenethylalcohol can be described as discriminating substances by means of GC/MS. With MCC-IMS, the results for 3-octanone and phenethylalcohol are concordant and additionally to GC/MS, ethanol and two further compounds (p_0642_1/p_683_1 and p_705_3) can be described. Isoamyl alcohol and cyclohexanone were not properly detectable with MCC-IMS. The major advantage of the MCC-IMS system is the feasibility of rapid analysis of complex gas mixtures without pre-concentration or preparation of samples and regardless of water vapour content in an online setup. Discrimination of fungi on genus level of the investigated germs by volatile metabolic profile and therefore detection of VOC is feasible. However, a further discrimination on species level for Candida species was not possible.     

In-vitro activities of terbinafine,itraconazole and amphotericin B against Aspergillus and Cladosporium species

The in vitro fungicidal and fungistatic activities of terbinafine were compared with those of itraconazole and amphotericin B against Aspergillus (n=63) and Cladosporium (n=21) isolates. The broth macrodilution modification of NCCLS reference method for filamentous fungi (M38-P) was used to assess the minimum inhibitory concentrations (MICs). Our data show that the in vitro activities of terbinafine were comparable to those of itraconazole and amphotericin B against the Aspergillus spp and Cladosporium strains tested. Despite strain-dependent variabilities, in general, itraconazole's activity was similar to that of amphotericin B against strains of Aspergillus and Cladosporium. Our data suggest that terbinafine may be useful in the treatment of Aspergillus and Cladosporium infections.     

Posaconazole: when and how? The clinician’s view

Posaconazole is the newest triazole antifungal agent available as an oral suspension with an extended spectrum of activity against Candida species, Aspergillus species, Cryptococcus neoformans, Zygomycetes and endemic fungi. Among posaconazole advantages are the relatively low potential of cross-resistance with other azoles, few drug interactions compared with other azoles and its activity against Zygomycetes. Randomised, double-blind trials have shown that posaconazole is effective for prophylaxis against invasive fungal infections (IFI), especially aspergillosis, in high-risk patients. Results of Phase III clinical trials and case/series reports indicate that posaconazole is effective in treating oesophageal candidiasis, including azole-refractory disease, and other IFI refractory to standard antifungal therapies. To date, posaconazole has appeared to be well tolerated even in long-term courses; it has an excellent safety profile with gastrointestinal disturbances being the most common adverse events reported. The dose of posaconazole is 200 mg three times daily for prophylaxis, 800 mg daily in two or four divided doses for the treatment of IFI and 100 mg daily (200 mg loading dose) for the treatment of oropharyngeal candidiasis. On the basis of early clinical experience, it appears that posaconazole will be a valuable aid in the management of life-threatening fungal infections.     

Prevalence of Aspergillus fumigatus and other fungal species in the sputum of adult patients with cystic fibrosis

Aspergillus fumigatus is often found in the respiratory tract secretions of patients with cystic fibrosis (CF), although the role of the fungus for progression of pulmonary disease remains unclear. This study aimed to investigate the frequency of A. fumigatus and other fungi in sputum of adult CF patients using different methods for culture and microscopy. Results from the analysis of 369 samples from 94 patients showed that A. fumigatus could be isolated in 45.7% of patients. Other moulds were rare, but the yeast Candida albicans was another frequent isolate, detected in 75.5% of patients. A comparison of different culture media showed no difference between a selective medium developed to specifically inhibit Pseudomonas aeruginosa and a standard fungal culture medium for growth of A. fumigatus, although both were more efficient for detection of fungi than other bacterial culture media. Fluorescent microscopy with calcofluor white was more sensitive for detection of fungal hyphae in undiluted sputum than standard methylene blue staining. This study shows that A. fumigatus and C. albicans have a high frequency in adult CF patients. Microbiological analysis should routinely include methods for specific identification of fungi to monitor for potential complications arising from fungal disease in these patients.     

In vitro activities of new and conventional antimycotics against fluconazole-susceptible and non-susceptible Brazilian Candida spp. isolates

The substantial increase in the rate of azole resistant Candida spp. yeast infections has become a serious treatment problem requiring new and more active antifungal agents. In this study, the in vitro activities of ravuconazole and albaconazole were compared with those of amphotericin B, flucytosine, itraconazole and fluconazole against 162 Brazilian isolates of Candida spp. from which 48 isolates had previously shown lower susceptibility or resistance to fluconazole. Ravuconazole susceptibility ranged from 84.6% (Candida albicans) to 100% for other species and albaconazole MIC(90) was < or =1.0 microg ml(-1) for all the species emphasising the potent activity of these triazoles. To our knowledge this is the first study evaluating the susceptibility of C. dubliniensis to albaconazole.     

Four cases of Candida albicans infections with isolates developing pink colonies on CHROMagar Candida plates

Candida albicans, the most commonly isolated yeast species, is typically identified by its green colony-colour on CHROMagar Candida plates. We here report four cases of Candida albicans infections, in which the initial identification was non-albicans isolates due to a clear pink colour of the colonies on CHROMagar Candida plates. However, classical phenotypic criteria, biochemical assimilation pattern and molecular characterisation identified all four isolates as C. albicans isolates.     

In vitro susceptibility-testing in Aspergillus species

Aspergillus species are the most common causes of invasive mould infections in immunocompromised patients. The introduction of new antifungal agents and recent reports of resistance emerging during treatment of Aspergillus infections have highlighted the need for in vitro susceptibility-testing. Various testing procedures have been proposed, including macrodilution and microdilution, agar diffusion, disc diffusion and Etest. At present, one of the most widely used assays is the M38-A reference method for filamentous fungi, published by the Clinical Laboratory Standard Institute and the Etest. Recently, the European Committee on Antimicrobial Susceptibility-testing (EUCAST) has charged its Antifungal Susceptibility-testing Subcommittee (AFST-EUCAST) with the preparation of new guidelines for in vitro susceptibility-testing of antifungals against Aspergillus spp. (EUCAST-AFST-ASPERGILLUS) defining breakpoints. This paper reviews the available methods for antifungal susceptibility-testing in Aspergillus spp. as well as the scant data regarding the clinical implications of in vitro testing.     

Molecular epidemiology of Candida species isolated from clinical specimens of intensive care unit patients

Epidemiological analysis of nosocomial Candida infections has gained importance due to an increase in these infections during the recent years. This study investigated the prevalence of clinical infections of Candida in anesthesiology intensive care unit patients, and ascertains the level of genetic diversity in the Candida species. A total of 70 Candida isolates, consisting of 42 Candida albicans, 16 Candida glabrata and 12 Candida tropicalis strains isolated from various clinical sites of infection of anesthesiology intensive care unit patients, were analysed. The susceptibility of the isolates against amphotericin B and fluconazole was determined by microdilution method according to Clinical and Laboratory Standards Institute M27-A2 standards. The strains were typed by random amplified polymorphic DNA (RAPD)-PCR using OPE-03, OPE-18, RP4-2 and AP50-1 primers. In the patients with Candida infections, most isolates exhibited different RAPD patterns. Only three C. albicans pairs isolated within a short time period had the same RAPD pattern. Most of the Candida infections in the anesthesiology intensive care unit of our hospital seem to be caused by endogenous strains. Exogenous spread of C. albicans infections occurred less frequently.     

Invasive fungal infections in the intensive care unit: a multicentre, prospective, observational study in Italy (2006-2008)

Critically ill patients admitted to intensive care units (ICU) are highly susceptible to healthcare-associated infections caused by fungi. A prospective sequential survey of invasive fungal infections was conducted from May 2006 to April 2008 in 38 ICUs of 27 Italian hospitals. A total of 384 fungal infections (318 invasive Candida infections, three cryptococcosis and 63 mould infections) were notified. The median rate of candidaemia was 10.08 per 1000 admissions. In 15% of cases, the infection was already present at the time of admission to ICU. Seventy-seven percent of Candida infections were diagnosed in surgical patients. Candida albicans was isolated in 60% of cases, Candida glabrata and Candida parapsilosis in 13%, each. Candida glabrata had the highest crude mortality rate (60%). Aspergillus infection was diagnosed in 32 medical and 25 surgical patients. The median rate was 6.31 per 1000 admissions. Corticosteroid treatment was the major host factor. Aspergillosis was demonstrated to be more severe than candidiasis as the crude mortality rate was significantly higher (63% vs. 46%), given an equal index of severity, Simplified Acute Physiology Score (SAPS-II). The present large nationwide survey points out the considerable morbidity and mortality of invasive fungal infections in surgical as well as medical patients in ICU.     

Activity and post antifungal effect of chlorpromazine and trifluopherazine against Aspergillus, Scedosporium and zygomycetes

The phenothiazine compounds chlorpromazine and trifluopherazine are antipsychotic agents that exhibit antimicrobial activity against bacteria, some protozoa and yeasts. Data of activity against filamentous fungi are lacking. The in vitro activity and postantifungal effect (PAFE) of chlorpromazine and trifluopherazine was determined against Aspergillus species, zygomycetes and Scedosporium species. In vitro susceptibility testing was performed with CLSI M38A and the PAFE was determined with previously established methods. Both drugs inhibited the growth of all fungi tested at concentrations of 16 to 64 microg ml(-1). For Aspergillus species the mean PAFE was 3.7 and 4.7 h; for zygomycetes, 3.1 and 3.4 h; for Scedosporium, 4.3 and 5.3 h for chlorpromazine and trifluoroperazine respectively. These are the first drugs shown to induce PAFE against Scedosporium. We show that phenothiazine compounds have in vitro antifungal activity and exhibit PAFE against a broad range of filamentous fungal pathogens. Although the exact mechanism of action is unknown, further studies are needed to explore the clinical usefulness of phenothiazine compounds.     

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.