As a result of fundamental changes in the International Code of Nomenclature on the use of separate names for sexual and asexual stages of fungi, generic names of many groups should be reconsidered. Members of the ECMM/ISHAM working group on Pseudallescheria/Scedosporium infections herein advocate a novel nomenclature for genera and species in Pseudallescheria , Scedosporium and allied taxa. The generic names Parascedosporium, Lomentospora, Petriella, Petriellopsis , and Scedosporium are proposed for a lineage within Microascaceae with mostly Scedosporium anamorphs producing slimy, annellidic conidia. Considering that Scedosporium has priority over Pseudallescheria and that Scedosporium prolificans is phylogenetically distinct from the other Scedosporium species, some name changes are proposed. Pseudallescheria minutispora and Petriellidium desertorum are renamed as Scedosporium minutisporum and S. desertorum , respectively. Scedosporium prolificans is renamed as Lomentospora prolificans .

Cystic fibrosis (CF) is the major genetic inherited disease in Caucasian populations. The respiratory tract of CF patients displays a sticky viscous mucus, which allows for the entrapment of airborne bacteria and fungal spores and provides a suitable environment for growth of microorganisms, including numerous yeast and filamentous fungal species. As a consequence, respiratory infections are the major cause of morbidity and mortality in this clinical context. Although bacteria remain the most common agents of these infections, fungal respiratory infections have emerged as an important cause of disease. Therefore, the International Society for Human and Animal Mycology (ISHAM) has launched a working group on Fungal respiratory infections in Cystic Fibrosis (Fri-CF) in October 2006, which was subsequently approved by the European Confederation of Medical Mycology (ECMM). Meetings of this working group, comprising both clinicians and mycologists involved in the follow-up of CF patients, as well as basic scientists interested in the fungal species involved, provided the opportunity to initiate collaborative works aimed to improve our knowledge on these infections to assist clinicians in patient management. The current review highlights the outcomes of some of these collaborative works in clinical surveillance, pathogenesis and treatment, giving special emphasis to standardization of culture procedures, improvement of species identification methods including the development of nonculture-based diagnostic methods, microbiome studies and identification of new biological markers, and the description of genotyping studies aiming to differentiate transient carriage and chronic colonization of the airways. The review also reports on the breakthrough in sequencing the genomes of the main Scedosporium species as basis for a better understanding of the pathogenic mechanisms of these fungi, and discusses treatment options of infections caused by multidrug resistant microorganisms, such as Scedosporium and Lomentospora species and members of the Rasamsonia argillacea species complex.

The primary aim of this study was to collect national epidemiological data on candidaemia and to determine the reporting time of species identification and antifungal susceptibility in clinical practice. During a 1-year period (March 2013 until February 2014), every first Candida isolate from each episode of candidaemia was included prospectively from 30 Belgian hospitals. Identification and susceptibility testing were performed according to local procedures and isolates were sent to the National Reference Center for Mycosis. Species identification was checked by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS) and internal transcribed spacer (ITS) sequencing in case no reliable identification was obtained by MALDI-TOF MS. Antifungal susceptibility testing was performed according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) methodology. A total of 355 isolates were retrieved from 338 patients. The mean incidence rate of candidaemia was 0.44 (range: 0.07 to 1.43) per 1000 admissions or 0.65 (range: 0.11 to 2.00) per 10,000 patient days. Candida albicans was most frequently found (50.4 %), followed by C. glabrata (27.3 %) and C. parapsilosis sensu lato (9.8 %). The overall resistance to fluconazole was 7.6 %, ranging from 3.9 % in C. albicans to 20.0 % in C. tropicalis. Only one C. glabrata isolate was resistant to the echinocandins. Four days after blood culture positivity, 99.7 % of the identifications and 90.3 % of the antifungal profiles were reported to the treating clinician. Candidaemia incidence rates differed up to 20-fold among Belgian hospitals; no clear factors explaining this difference were identified. The overall antifungal resistance rates were low but high azole resistance rates were recorded in C. tropicalis.

What do you need to make a Higgs boson, or travel to the event horizon of a black hole. Answers could be lurking in many a humble physics lab

Arthroderma benhamiae is a zoophilic dermatophyte belonging to the Trichophyton mentagrophytes species complex. Here, a population of A. benhamiae wild strains from the same geographical area (Switzerland) was studied by comparing their morphology, assessing their molecular variability using internal transcribed spacer (ITS) and 28S rRNA gene sequencing, and evaluating their interfertility. Sequencing of the ITS region and of part of the 28S rRNA gene revealed the existence of two infraspecific groups with markedly different colony phenotypes: white (group I) and yellow (group II), respectively. For all strains, the results of mating type identification by PCR, using HMG (high-mobility group) and α-box genes in the mating type locus as targets, were in total accordance with the results of mating type identification by strain confrontation experiments. White-phenotype strains were of mating type + (mt+) or mating type - (mt-), whilst yellow-phenotype strains were all mt-. White and yellow strains were found to produce fertile cleistothecia after mating with A. benhamiae reference tester strains, which belonged to a third group intermediate between groups I and II. However, no interfertility was observed between yellow strains and white strains of mt+. A significant result was that white strains of mt- were able to mate and produce fertile cleistothecia with the white A. benhamiae strain CBS 112371 (mt+), the genome of which has recently been sequenced and annotated. This finding should offer new tools for investigating the biology and genetics of dermatophytes using wild-type strains.

A 54-year-old immunocompetent man presented with an infrapatellar bursitis caused by Prototheca wickerhamii. Because of clinical and microbiological relapse two weeks after bursectomy, six weekly injections of 5 mg of conventional amphotericin B were chosen for intrabursal treatment. Four months after completion of the treatment, the patient remains cured.

The black yeast Exophiala dermatitidis is a frequent agent of colonization of the lungs of patients with cystic fibrosis (CF). A total of 71 clinical isolates of Exophiala from 13 patients were identified at the species level by sequencing the internal transcribed spacer (ITS) regions 1 and 2 of the rDNA genes and typed by random amplification of polymorphic DNA (RAPD), using two different primers, BG-2 and ERIC-1. In vitro susceptibility of these isolates to some systemic antifungal drugs was investigated using the CLSI method. Almost all the isolates were identified as E. dermatitidis, but long-term colonization with the closely related species E. phaeomuriformis was observed in one patient. No clustering was found according to the geographical origin of the isolates, the isolation date or the antifungal susceptibility. Variations were seen in the susceptibility of studied isolates to antifungals but most of them exhibited low susceptibility to amphotericin B and although some patients were successively colonized by two distinct genotypes, most of the isolates were distributed in patient-specific clusters. This phenomenon may be due to genomic variations of E. dermatitidis in the lung environment of CF patients. These results are typical of colonization of the airways of patients by a poorly distributed environmental fungus, which occupies particular reservoirs that need to be defined.

Background Surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) was applied to analyze the protein profiles in both somatic and metabolic extracts of Aspergillus species. The study was carried out on some Aspergillus species within the Fumigati section ( Aspergillus fumigatus wild-types and natural abnormally pigmented mutants, and Aspergillus lentulus ). The aim was to validate whether mass spectrometry protein profiles can be used as specific signatures to discriminate different Aspergillus species or even mutants within the same species. Results The growth conditions and the SELDI-TOF parameters were determined to generate characteristic protein profiles of somatic and metabolic extracts of Aspergillus fumigatus strains using five different ProteinChips ® , eight growth conditions combining two temperatures, two media and two oxygenation conditions. Nine strains were investigated: three wild-types and four natural abnormally pigmented mutant strains of A. fumigatus and two strains of A. lentulus . A total of 242 fungal extracts were prepared. The spectra obtained are protein signatures linked to the physiological states of fungal strains depending on culture conditions. The best resolutions were obtained using the chromatographic surfaces CM10, NP20 and H50 with fractions of fungi grown on modified Sabouraud medium at 37°C in static condition. Under these conditions, the SELDI-TOF analysis allowed A. fumigatus and A. lentulus strains to be grouped into distinct clusters. Conclusions SELDI-TOF analysis distinguishes A. fumigatus from A. lentulus strains and moreover, permits separate clusters of natural abnormally pigmented A. fumigatus strains to be obtained. In addition, this methodology allowed us to point out fungal components specifically produced by a wild-type strain or natural mutants. It offers attractive potential for further studies of the Aspergillus biology or pathogenesis.

Dermatophytes are keratinophilic fungi that can be pathogenic for humans and animals by infecting the stratum corneum, nails, claws or hair. The first infection step consists of adherence of arthroconidia to the stratum corneum. The mechanisms and the kinetics of adherence have been investigated using different in vitro and ex vivo experimental models, most notably showing the role of a secreted serine protease from Microsporum canis in fungal adherence to feline corneocytes. After germination of the arthroconidia, dermatophytes invade keratinised structures that have to be digested into short peptides and amino acids to be assimilated. Although many proteases, including keratinolytic ones, have been characterised, the understanding of dermatophyte invasion mechanisms remains speculative. To date, research on mechanisms of dermatophyte infection focused mainly on both secreted endoproteases and exoproteases, but their precise role in both fungal adherence and skin invasion should be further explored.

Introduction: Prognosis of cystic fibrosis (CF) essentially depends on impairment of the lung function. While considerable attention therefore has been paid over recent decades to the prevention and treatment of bacterial respiratory infections, leading to a marked increase in life expectancy of the patients, prevalence of colonization of the airways by filamentous fungi or yeasts, sometimes leading to true respiratory infections, has been regularly increasing. However, recently, Borman et al. (2010) reported wide variations in the range of the causative fungal pathogens and in their prevalence, related at least to a lack of standardization in the methods used to detect these microorganisms. Here, our aims are: to analyze the impact of the culture conditions used on the detection of specific fungal pathogens throughout the French multicenter experience, and to discuss the methods used in various European or Australian laboratories, in order to carry out an international prospective study that will make possible a standardized protocol for efficient detection of the whole fungal biota that can be encountered in respiratory secretions of CF patients. Results: First, from January 2007 to the end of 2009, a multicenter study was conducted in France encompassing 7 university or general hospitals which agreed to use the same procedure for mycological analysis of sputum saples from CF patients, including prior digestion of the sample with dithiolbutane and inoculation of the digested sample on 6 semi- selective agar-based media (MucoFong study – PHRC1902). Data obtained during one year were analyzed using the CHAID (Chi-squared Automatic Interaction Detector) method, which is a statistical approach able to give best association of media to detect a specific pathogen. CHAID is a type of decision tree technique, based upon adjusted significance testing that we apply to our data in order to define the best set of semi-selective media able to isolate 99.99% of the fungal pathogens that were detected in our CF population. Second, the major data of the international survey will be presented, and discussed with the purpose of developing a standardized approach for mycological examination of respiratory secretions from CF patients. Conclusion: Defining the optimal method for mycological analysis of the fungal components of CF lungs microbiome through a large international study is becoming a major requirement. This will make possible not only to analyze the role of some rare filamentous species in CF exacerbation or the existence of geographic variations in the fungal species that colonize the airways, but also to study the complexity of the CF lung microbiome as well as its dynamics.

In this study, three different rat hybridoma cell lines secreting monoclonal antibodies (mAbs) recognizing the spores from Alternaria alternata, a plant pathogenic fungus, contaminant of food products and important cause of both allergic rhinitis and asthma, have been characterized. These three mAbs are all of IgM isotype. Two antibodies, A1 and F10, were cross-reactive antibodies recognizing spores from Alternaria, Cladosporium, Penicillium, Aspergillus and Stachybotrys genera, but not the yeasts Saccharomyces cerevisiae or Candida albicans. Competitive and sandwich assays demonstrated that these two mAbs were directed against the same or very close repetitive(s) epitope(s). A1-based sandwich ELISA efficiently detected this epitope in various mould (but not yeast)-soluble extracts prepared from strains grown in the laboratory. Moreover, this A1-based sandwich ELISA detected its cognate epitope in air and dust samples obtained from dwellings. The third antibody, E5, recognized only the spores of Alternaria and the phylogenetically very close Ulocladium botrytis. This E5 antibody is directed against a repetitive epitope found in Alternaria and Ulocladium laboratory extracts and can be used in a sandwich assay for the quantification of these moulds. Therefore, E5 antibody is a promising tool for the development of Alternaria–Ulocladium–specific immunoassays, while A1 and F10 could be interesting tools for the quantification of the total mould biomass.

A novel dermatophyte species is described in the Microsporum cookei clade. It differs significantly from known taxa in the two molecular markers analyzed, i.e., ITS and partial β-tubulin (BT2). Morphologically the species was characterized by smooth- or only slightly rough-walled conidia, but isolates rapidly became pleomorphic with sparse, smooth- and thick-walled macroconidia in addition to microconidia. A teleomorph was found after mating.

Research on orphan diseases has been boosted enormously over the last decade with the event of electronic communication. This has enabled the implementation of international networks providing research groups with sufficient critical mass for epidemiological studies. An example of such a success story is without doubt the knowledge on Scedosporium and its teleomorph Pseudallescheria. Although already known from human infections since the late 19th century, these fungi had long been regarded either as clinically insignificant, or as anecdotal. Today the species are listed among the major groups of filamentous opportunists.1,2 First attempts to unite researchers and clinicians were made by the Spanish Study Group on Scedosporium prolificans. In 2002, a Europe-wide group was founded under the umbrella of the European Confederation of Medical Mycology (ECMM). As similar initiatives were undertaken in Australia by the Australian Scedosporium Study Group (AUSCEDO), the two groups were internationalized under the auspices of the International Society of Human and Animal Mycology (ISHAM). Main objective of the Working Group Pseudallescheria/Scedosporium Infections was to gain insight into the epidemiology and genetic variability of these fungi and to provide data on possible sources of contamination and routes of infection. The taxonomy of the fungi had been revolutionised by the application of molecular methods, particularly through the papers of Gilgado et al.[3–5] The classical species Pseudallescheria boydii was subdivided into numerous species, several of which were indistinguishable by phenotypic characteristics that had been in use until recently. A complicating factor is the different degree of inbreeding and clonality between species, leaving doubt whether all genealogically separated entities can be referred to as species in the classical sense.6 In addition, it remains questionable whether distinction of all entities is clinically meaningful. But at least there is a wide consensus that Pseudallescheria is a species complex rather than a single species. Species have limited molecular heterogeneity and comprise limited numbers of haplotypes. A number of molecular techniques for diagnostics and detection are currently being developed using genes that have been suitable for identification of species. The widely used rDNA internal transcribed spacer (ITS) region of rDNA is suitable for the majority of clearly distinct taxa,7,8 whereas molecular siblings are separable by different loci in the β-tubulin gene.3,4 Scedosporium species are opportunists and thus understanding of their behaviour in human tissue can be reached only via knowledge of their environmental habitat. Kaltseis et al. [9] noted that Scedosporium species are positively associated with human-derived, industrial and agricultural pollution. Comparing the frequency of species from the environment with the distribution of species involved in human infection, the authors supposed significant difference in virulence between species. Virulence is concentrated in two locations in the phylogeny of Microascales with Scedosporium-like appearance, viz. in the Pseudallescheria boydii complex discussed above, and in Scedosporium prolificans.10 These fungi primarily cause subcutaneous infections in healthy individuals, or deep, occasionally disseminated infections in debilitated patients. A remarkable, newly recognised clinical syndrome is the near-drowning encephalitis, a delayed infection of the brain after aspiration of polluted water resulting in temporary coma.11,12 With improved isolation and detection techniques13–16Scedosporium species have also become recognised as common colonisers of the airways of patients with cystic fibrosis.17 The direct clinical significance of this finding is still unclear,18 but infection may be regarded as a contraindication for lung transplantation,19,20 the ultimate therapy for CF patients. Scedosporium infections are notoriously difficult to treat due to their limited susceptibility to most commonly used systemic antifungals. Species share this property with the Scopulariopsis agents of cutaneous infections, which belong to the same order, Microascales. In the filamentous ascomycetes such recalcitrance to therapy is matched only by the order Hypocreales, containing the genera Acremonium, Fusarium and Trichoderma. Scedosporium prolificans belongs to the fungi with the highest degree of resistance to antifungals known. Reasonable results have been obtained with combination therapy using voriconazole and terbinafin,21,22 but in general mortality rates in disseminated infections by this fungus rise until up to 87.5%.23 Infections caused by species of the P. boydii complex have proved less difficult to treat, with voriconazole being the drug of choice.24,25 Strains are currently identifiable with the sequence base available at http://www.scedosporium-ecmm.com. Species concepts applied in this database have been verified by multilocus analysis including several gene loci (ITS, BT2, TUB) and AFLP profiles, and taxonomy has been anchored by the inclusion of type strains. The database is divided up between clinical and environmental strains (Fig. 1) because metadata for the two categories are very different, but the identification procedure is identical. ITS and the BT2 and TUB loci of β-tubulin are sufficient for reliable identification. At the University of Sydney Westmead Hospital, a database for multilocus sequence typing applying six genetic loci for Scedosporium aurantiacum was developed and is accessible at http://mlst.mycologylab.org (A. Harun & W. Meyer, unpublished data). Clinical data are automatically transmitted to the Fungiscope database, where tools for epidemiological analysis are being installed. Deposition of live material is recommended in one of the recognised culture collections joining the project (Fig. 1), whereby the Belgian Coordinated Collection of Microorganisms at the Scientific Institute of Public Health (Brussels) serves as a prime depository for environmental strains. Strains are available to members of the Working Group if permission from the depositor is granted. A taxonomic database is available through MycoBank (http://www.mycobank.org), while a nearly complete collection of clinical papers published before 2006 is available on the ISHAM website (http://www.isham.org). Note that there is no link to GenBank, as this database is not updated according to taxonomic developments and sequences are not verified with ex-type materials. Flow diagram of data storage of clinical and environmental strains. The cooperating Working Groups have thus provided a basic infrastructure that is essential for the growth of knowledge on Pseudallescheria and Scedosporium. We offer access to a broad range of information on these emerging fungal opportunists, which should lead to appropriate and effective therapy. Clearly, the success of this endeavour depends on the ongoing activity of its supporters. Readers of this special issue are cordially invited to contribute to the network. The present special issue stems from presentations given at the PSI workshop held in Bonn, Germany, 6−8 May, 2010. The authors have no conflict of interests to declare.

The objective of this prospective study was to assess the prevalence of Exophiala dermatitidis in respiratory secretions of patients with cystic fibrosis (CF) and to identify risk factors for its presence. The results of all cultures performed over a 2-year period in non lung-transplant patients in our CF clinic were included in the study. Samples consisted of sputum (whenever possible) or deep pharyngeal aspirate after a session of physiotherapy. Specimens were inoculated onto Sabouraud gentamicin-chloramphenicol agar (SGCA) medium (Becton-Dickinson) and incubated at 35°C for 2 days and then at ambient temperature (15-25°C) for 3 weeks. The whole study group included 154 patients (mean age ± SD: 18.5 y ± 11.69). E. dermatitidis was isolated from 58 specimens (2.8%) of nine patients (5.8%) out of total of 2065 cultures prepared during the study period. All E. dermatitidis culture-positive patients were pancreatic insufficient and ≥12 y of age. Almost all (8/9) were homozygous for the F508 del mutation. Aspergillus fumigatus colonization and genotype seemed to be predisposing factors. No other significant characteristic was identified in this group, either in terms of predominant bacterial pathogen or treatment. A distinct comparative study performed over 3 months in our laboratory revealed that the use of SGCA yielded identical isolation rates of E. dermatitidis as erythritol-chloramphenicol agar (ECA).

Isolates of the Trichophyton mentagrophytes complex vary phenotypically. Whether the closely related zoophilic and anthropophilic anamorphs currently associated with Arthroderma vanbreuseghemii have to be considered as members of the same biological species remains an open question. In order to better delineate species in the T. mentagrophytes complex, we performed a mating analysis of freshly collected isolates from humans and animals with A. benhamiae and A. vanbreuseghemii reference strains, in comparison to internal transcribed spacer (ITS) and 28S rDNA sequencing. Mating experiments as well as ITS and 28S sequencing unambiguously allowed the distinction of A. benhamiae and A. vanbreuseghemii. We have also shown that all the isolates from tinea pedis and tinea unguium identified as T. interdigitale based on ITS sequences mated with A. vanbreuseghemii tester strains, but had lost their ability to give fertile cleistothecia. Therefore, T. interdigitale has to be considered as a humanized species derived from the sexual relative A. vanbreuseghemii.