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Wood is an important construction material, but when used incorrectly it can be subjected to deterioration by wood-destroying fungi. The brown rot producing dry rot fungus (Serpula lacrymans) is by far the most dangerous wood-destroying fungus in Europe. In the present publication, 645 fungal samples from damaged wood in the federal state of Styria (Austria) were examined and recorded by isolation date, geographical location, species identification of the wood-destroying fungus, location of damage, construction method, and age and type of building. In Styria, Serpula spp. accounted for 61.5% of damages, followed by Antrodia spp. (10.7%) and the genera Gloeophyllum (8.2%), Coniophora (3.9%) and Donkioporia (1.1%). Properties in the area of the Styrian capital Graz and old buildings were more often infested by wood-destroying fungi than houses in the rural area and new constructions.

BACKGROUND Candida-associated infections put a significant burden on western healthcare systems. Development of (multi-)resistant fungi can become untreatable and threaten especially vulnerable target groups, such as the immunocompromised. OBJECTIVES We assessed antifungal susceptibility and explored possible influence factors of clinical Candida isolates collected from Austrian hospitals between 2007 and 2016. METHODS Thousand three hundred and sixty clinical Candida spp. isolated from blood cultures were subjected to antifungal susceptibility testing (AFST) in a liquid-handling aided continuous microdilution assay. We tested against fluconazole, voriconazole, posaconazole, itraconazole, isavuconazole, anidulafungin, caspofungin and micafungin according to EUCAST with additional recording of growth curves. We performed rigid quality control on each assay via growth curve assessment and included two standard reference strains. Minimal inhibitory concentrations (MIC) were quantified according to EUCAST guideline E.DEF 7.3.1, and susceptibility was evaluated using EUCAST clinical breakpoints. RESULTS The isolate collection consisted of Candida albicans (59%), C. glabrata (19%), C. parapsilosis (9%), C. tropicalis (5%) and C. krusei (3%) and few other Candida species and fungi (5%). During the observed time period, species abundance and antifungal resistance rates remained constant. Multi-resistance was rare and we found no single isolate which was resistant to both azoles and echinocandins. Within the antifungal resistance profile of our strain collection, we observed clusters along species boundaries. CONCLUSIONS Over the last decade, the distribution of Candida species and its level of antifungal resistance remained constant in Austria. Our data compare well with other European countries. Principal component analysis of the susceptibility profile of this collection revealed species-specific clusters and substantial intra-species variation, especially for C. glabrata.

An account is provided of the world’s ten most feared fungi. Within areas of interest, we have organized the entries in the order of concern. We put four human pathogens first as this is of concern to most people. This is followed by fungi producing mycotoxins that are highly harmful for humans; Aspergillus flavus, the main producer of aflatoxins, was used as an example. Problems due to indoor air fungi may also directly affect our health and we use Stachybotrys chartarum as an example. Not everyone collects and eats edible mushrooms. However, fatalities caused by mushroom intoxications often make news headlines and therefore we include one of the most poisonous of all mushrooms, Amanita phalloides, as an example. We then move on to the fungi that damage our dwellings causing serious anxiety by rotting our timber structures and flooring. Serpula lacrymans, which causes dry rot is an excellent example. The next example serves to represent all plant and forest pathogens. Here we chose Austropuccinia psidii as it is causing devastating effects in Australia and will probably do likewise in New Zealand. Finally, we chose an important amphibian pathogen which is causing serious declines in the numbers of frogs and other amphibians worldwide. Although we target the top ten most feared fungi, numerous others are causing serious concern to human health, plant production, forestry, other animals and our factories and dwellings. By highlighting ten feared fungi as an example, we aim to promote public awareness of the cost and importance of fungi.

Die von der Gesellschaft fur Hygiene, Umweltmedizin und Praventivmedizin (GHUP) federfuhrend erstellte Leitlinie „Medizinisch klinische Diagnostik bei Schimmelpilzexposition in Innenraumen“ ist Gegenstand des vorliegenden Abschnitts. Schimmelpilzwachstum im Innenraum ist als ein potenzielles Gesundheitsrisiko zu betrachten, auch ohne dass ein quantitativer und/oder kausaler Zusammenhang zwischen dem Vorkommen einzelner Arten und Gesundheitsbeschwerden gesichert werden kann. Abgesehen von der Allergischen Bronchopulmonalen Aspergillose (ABPA) und den durch Schimmelpilze kausal verursachten Mykosen, liegen lediglich ausreichende Evidenzen fur folgende Assoziationen von Feuchte-/Schimmelpilzschaden und unterschiedlichen Gesundheitseffekten vor: allergische Atemwegserkrankungen, Asthma (Manifestation, Progression, Exazerbation), allergische Rhinitis, Exogen Allergische Alveolitis, Begunstigung von Atemwegsinfekten/Bronchitis. Dabei ist das sensibilisierende Potenzial von Schimmelpilzen im Vergleich zu anderen Umweltallergenen deutlich geringer einzuschatzen. Aktuelle Studien zeigen europaweit eine vergleichsweise geringe Sensibilisierungspravalenz von 3–10 % gemessen an der Gesamtbevolkerung. Eingeschrankte oder vermutete Evidenz fur eine Assoziation liegt vor hinsichtlich „mucous membrane irritation“ und Atopischen Ekzems (Manifestation, Progression, Exazerbation). Inadaquate oder unzureichende Evidenz fur eine Assoziation liegt vor fur COPD, akute idiopathische pulmonale Hamorrhagie bei Kindern, Rheuma/Arthritis, Sarkoidose und Krebserkrankungen. Das Infektionsrisiko von den in Innenraumen regelmasig vorkommenden Schimmelpilzarten ist fur gesunde Personen gering, die meisten Arten sind in die Risikogruppe 1 und wenige in 2 (Aspergillus fumigatus, A. flavus) der Biostoffverordnung eingestuft. Nur Schimmelpilze, die potenziell in der Lage sind, Toxine zu bilden, kommen als Ausloser einer Intoxikation in Betracht. Ob im Einzelfall eine Toxinbildung im Innenraum stattfindet, entscheiden die Umgebungs- und Wachstumsbedingungen und hier vor allem das Substrat. Von Geruchswirkungen und/oder Befindlichkeitsstorungen kann bei Feuchte-/Schimmelpilzschaden im Innenraum grundsatzlich jeder betroffen sein. Hierbei handelt es sich nicht um eine Gesundheitsgefahrdung. Pradisponierende Faktoren fur Geruchswirkungen konnen genetische und hormonelle Einflusse, Pragung, Kontext und Adaptationseffekte sein. Pradisponierende Faktoren fur Befindlichkeitsstorungen konnen Umweltbesorgnisse, -angste, -konditionierungen und -attributionen sowie eine Vielzahl von Erkrankungen sein. Besonders zu schutzende Risikogruppen bezuglich eines Infektionsrisikos sind Personen unter Immunsuppression nach der Einteilung der Kommission fur Krankenhaushygiene und Infektionspravention (KRINKO) beim Robert Koch-Institut (RKI) und Personen mit Mukoviszidose (Zystischer Fibrose), bezuglich eines allergischen Risikos Personen mit Mukoviszidose (Zystischer Fibrose) und Personen mit Asthma bronchiale. Die rationale Diagnostik beinhaltet die Anamnese, eine korperliche Untersuchung, eine konventionelle Allergiediagnostik einschlieslich gegebenenfalls Provokationstests, vereinzelt sind auch zellulare Testsysteme indiziert. Zum Vorgehen bei Schimmelpilzinfektionen wird auf die angemeldete AWMF-Leitlinie Diagnose und Therapie invasiver Aspergillus-Infektionen verwiesen. Hinsichtlich Mykotoxine existieren zurzeit keine brauchbaren und validierten Testverfahren, die in der klinischen Diagnostik eingesetzt werden konnten. Praventivmedizinisch ist wichtig, dass Schimmelpilzbefall in relevantem Ausmas aus Vorsorgegrunden nicht toleriert werden darf. Zur Beurteilung des Schadensausmases und zum Vorgehen wird auf den „Schimmelpilzleitfaden“ des Umweltbundesamtes verwiesen.

This article is an abridged version of the AWMF mould guideline “Medical clinical diagnostics of indoor mould exposure” presented in April 2016 by the German Society of Hygiene, Environmental Medicine and Preventive Medicine (Gesellschaft für Hygiene, Umweltmedizin und Präventivmedizin, GHUP), in collaboration with the above-mentioned scientific medical societies, German and Austrian societies, medical associations and experts. Indoor mould growth is a potential health risk, even if a quantitative and/or causal relationship between the occurrence of individual mould species and health problems has yet to be established. Apart from allergic bronchopulmonary aspergillosis (ABPA) and mould-caused mycoses, only sufficient evidence for an association between moisture/mould damage and the following health effects has been established: allergic respiratory disease, asthma (manifestation, progression and exacerbation), allergic rhinitis, hypersensitivity pneumonitis (extrinsic allergic alveolitis), and increased likelihood of respiratory infections/bronchitis. In this context the sensitizing potential of moulds is obviously low compared to other environmental allergens. Recent studies show a comparatively low sensitizing prevalence of 3–10% in the general population across Europe. Limited or suspected evidence for an association exist with respect to mucous membrane irritation and atopic eczema (manifestation, progression and exacerbation). Inadequate or insufficient evidence for an association exist for chronic obstructive pulmonary disease, acute idiopathic pulmonary hemorrhage in children, rheumatism/arthritis, sarcoidosis and cancer. The risk of infection posed by moulds regularly occurring indoors is low for healthy persons; most species are in risk group 1 and a few in risk group 2 (Aspergillus fumigatus, A. flavus) of the German Biological Agents Act (Biostoffverordnung). Only moulds that are potentially able to form toxins can be triggers of toxic reactions. Whether or not toxin formation occurs in individual cases is determined by environmental and growth conditions, above all the substrate. In the case of indoor moisture/mould damage, everyone can be affected by odour effects and/or mood disorders. However, this is not a health hazard. Predisposing factors for odour effects can include genetic and hormonal influences, imprinting, context and adaptation effects. Predisposing factors for mood disorders may include environmental concerns, anxiety, condition, and attribution, as well as various diseases. Risk groups to be protected particularly with regard to an infection risk are persons on immunosuppression according to the classification of the German Commission for Hospital Hygiene and Infection Prevention (Kommission für Krankenhaushygiene und Infektionsprävention, KRINKO) at the Robert Koch- Institute (RKI) and persons with cystic fibrosis (mucoviscidosis); with regard to an allergic risk, persons with cystic fibrosis (mucoviscidosis) and patients with bronchial asthma should be protected. The rational diagnostics include the medical history, physical examination, and conventional allergy diagnostics including provocation tests if necessary; sometimes cellular test systems are indicated. In the case of mould infections the reader is referred to the AWMF guideline “Diagnosis and Therapy of Invasive Aspergillus Infections”. With regard to mycotoxins, there are currently no useful and validated test procedures for clinical diagnostics. From a preventive medicine standpoint it is important that indoor mould infestation in relevant dimension cannot be tolerated for precautionary reasons. With regard to evaluating the extent of damage and selecting a remedial procedure, the reader is referred to the revised version of the mould guideline issued by the German Federal Environment Agency (Umweltbundesamt, UBA).

© 2018 Background: Fungi are increasingly recognized for their potential role in contributing to pulmonary damage in Cystic Fibrosis (CF). We therefore designed a prospective international study aimed at (i) determining the prevalence of fungi isolated from sputum samples collected from a large CF population, (ii) comparing the performance of different media used for fungal culture, and (iii) proposing a standardized protocol suitable for CF routine microbiology. Methods: An international, consensually designed prospective study was set up (https://www.ecfs.eu/special-projects/mucofong-international-project). All centers worked according to the same protocol approved by Lille Ethical Committee. CF sputa were inoculated onto eight semi-selective media incubated at 37 °C or 25 °C–30 °C for 15 days, and inspected twice weekly for fungal growth. Results: A total of 469 sputa were collected from patients at 18 European and one Australian CF centers. Positive cultures for fungal growth were significantly associated with patient ages. Aspergillus fumigatus was the most frequently isolated mold. We identified a growing European North-to-South gradient of Scedosporium prevalence, while yeasts, Aspergillus section Fumigati, Cladosporium and Penicillium were significantly more prevalent in the Northern regions. Conclusions: According to the CHi-squared Automatic Interaction Detector method, we propose a consensual protocol based on two media (YPDA or Sabouraud medium, and B(+) medium) to detect the main opportunistic molds in CF context; the use of an additional medium being recommended according to the patient's clinical status. This standardized protocol allows us to have an accurate overview of the respiratory mycobiome on the culturomic side in CF. ispartof: JOURNAL OF CYSTIC FIBROSIS vol:18 issue:2 pages:212-220 ispartof: location:Netherlands status: published

The objective of this study was to investigate the prognostic potential of 1,3-beta-d-glucan (BDG) testing in bronchoalveolar lavage fluid (BALF) samples.A total of 300 BALF samples from 252 patients were investigated for BDG (Fungitell(®) assay). Prognostic potential of BALF BDG was evaluated by using: i.) Kaplan-Meier analysis, and ii.) multivariable Cox hazard regression analyses.BALF BDG levels were found to be significantly higher in samples with Candida spp. colonization (p 0.001). A total of 61/252 patients (24.2%) died within 90-days of BALF sampling (18.1% of patients with BALF BDG200 pg/mL, 32.4% with BALF BDG ≥200 pg/mL). Kaplan-Meier analysis revealed that overall cumulative 90-day mortality was significantly higher in those with BALF BDG levels ≥200 pg/mL when compared to those with levels200 pg/mL (log-rank p = 0.006, Breslow p = 0.005 and Tarone-Ware p = 0.005). The multivariable Cox regression analysis showed that BALF BDG levels were a strong predictor of 90-day overall mortality, with a hazard ratio of 1.048 (per 100 pg/mL increase of BALF BDG).False positive BALF BDG results in the presence of Candida spp. colonization of the lower respiratory tract may explain the limited diagnostic potential of BALF BDG testing. In contrast, prognostic potential of BALF BDG may be promising.

The achievement of a better life for cystic fibrosis (CF) patients is mainly caused by a better management and infection control over the last three decades. Herein, we want to summarize the cornerstones for an effective management of CF patients and to give an overview of the knowledge about the fungal epidemiology in this clinical context in Europe. Data from a retrospective analysis encompassing 66,616 samples from 3235 CF patients followed-up in 9 CF centers from different European countries are shown.

International audience; 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.

The emerging pathogen Candida auris is isolated mostly from hospitalized patients and often shows multidrug resistance. We report on the isolation of this yeast in Austria from an outpatient's auditory canal. The isolate showed good susceptibility against antifungals except for echinocandins; the patient was treated successfully with topical administration of nystatin.

Pu-erh tea originates from the province of Yunnan in south-western China. As this tea is produced by so called Aspergillus post-fermentation the question arises which molds and mycotoxins may be found in this tea. In total 36 samples of Pu-erh tea were investigated for their content of filamentous fungi and the mycotoxins aflatoxins B1, B2, G1, and G2, fumonisins B1, B2, and B3, and ochratoxin A. Fungi were isolated from all samples in a concentration of 1.0×10(1) to 2.6×10(6) colony forming units (cfu)/g tea, all together 19 fungal genera and 31 species were identified. The most prevalent species were Aspergillus acidus and Aspergillus fumigatus, followed by Zygomycetes and Penicillium species. Aflatoxins and fumonisins were not found in the samples investigated, ochratoxin A was detected in 4 of 36 teas (11.1%).

The ambient air consists not only of gases but also of bioaerosols and particulate matter. The concentrations of particulate matter in relation to the culturable microorganisms in the urban ambient air and their dependence on air temperature and relative humidity were investigated. The seasonal distribution of particles sizes, the concentrations of aerobic mesophilic bacteria and xerophilic fungi in the air were evaluated. Moreover, the identification of the fungal genera Cladosporium, Aspergillus and Penicillium were conducted. Within one year at 177 days particle and microorganism concentrations in the ambient air were recorded in the city centre of Graz/Austria. The results show that the concentrations of fine particles and coarse particles were the highest in winter and decreased continuously to a minimum in the summer months depending on temperature and air humidity. The concentrations of xerophilic fungi showed no correlation to the different particle concentrations. The spore concentrations of Cladosporium spp. showed the same results of xerophilic fungi whereas the genera Penicillium and Aspergillus increased with the increase of fine particles. The concentrations of mesophilic bacteria were positively correlated with all particle counts. The maximum mesophilic bacteria concentrations were found in the winter months. Further studies are required to evaluate the concentrations of specific microorganisms in the natural environment in relation to the particulate matter.

In April 2016, the German Society of Hygiene, Environmental Medicine and Preventative Medicine (Gesellschaft fur Hygiene, Umweltmedizin und Praventivmedizin (GHUP)) together with other scientific medical societies, German and Austrian medical societies, physician unions and experts has provided an AWMF (Association of the Scientific Medical Societies) guideline 'Medical diagnostics for indoor mold exposure'. This guideline shall help physicians to advise and treat patients exposed indoors to mold. Indoor mold growth is a potential health risk, even without a quantitative and/or causal association between the occurrence of individual mold species and health effects. Apart from the allergic bronchopulmonary aspergillosis (ABPA) and the mycoses caused by mold, there is only sufficient evidence for the following associations between moisture/mold damages and different health effects: Allergic respiratory diseases, asthma (manifestation, progression, exacerbation), allergic rhinitis, exogenous allergic alveolitis and respiratory tract infections/bronchitis. In comparison to other environmental allergens, the sensitizing potential of molds is estimated to be low. Recent studies show a prevalence of sensitization of 3-10% in the total population of Europe. The evidence for associations to mucous membrane irritation and atopic eczema (manifestation, progression, exacerbation) is classified as limited or suspected. Inadequate or insufficient evidence for an association is given for COPD, acute idiopathic pulmonary hemorrhage in children, rheumatism/arthritis, sarcoidosis, and cancer. The risk of infections from indoor molds is low for healthy individuals. Only molds that are capable to form toxins can cause intoxications. The environmental and growth conditions and especially the substrate determine whether toxin formation occurs, but indoor air concentrations are always very low. In the case of indoor moisture/mold damages, everyone can be affected by odor effects and/or impairment of well-being. Predisposing factors for odor effects can be given by genetic and hormonal influences, imprinting, context and adaptation effects. Predisposing factors for impairment of well-being are environmental concerns, anxieties, conditioning and attributions as well as a variety of diseases. Risk groups that must be protected are patients with immunosuppression and with mucoviscidosis (cystic fibrosis) with regard to infections and individuals with mucoviscidosis and asthma with regard to allergies. If an association between mold exposure and health effects is suspected, the medical diagnosis includes medical history, physical examination, conventional allergy diagnosis, and if indicated, provocation tests. For the treatment of mold infections, it is referred to the AWMF guidelines for diagnosis and treatment of invasive Aspergillus infections. Regarding mycotoxins, there are currently no validated test methods that could be used in clinical diagnostics. From the perspective of preventive medicine, it is important that mold damages cannot be tolerated in indoor environments.

The aim of this study was to determine the prevalence of invasive aspergillosis (IA) in patients with liver cirrhosis and the performance of serum galactomannan (GM) screening. Patients with decompensated liver cirrhosis and patients with compensated liver cirrhosis presenting with fever and/or respiratory symptoms were prospectively enrolled. All patients were screened by serum GM twice weekly irrespective of clinical signs and symptoms. Positive serum GM triggered work-up consisting of chest computed tomography and in case of pathological findings bronchoscopy. 150 patients were included in the study. Two (1.3%) had probable, one (0.7%) had possible, and 147 (98%) had no evidence of IA. Both patients with probable IA had compensated liver cirrhosis. Sensitivity for serum GM screening for probable versus no IA was 0.5 (95% CI, 0.09-0.91), specificity 0.97 (95% CI: 0.92-0.99), negative predictive value 0.99 (95% CI, 0.96-0.99) and positive predictive value (PPV) 0.17 (95% CI, 0.01-0.64). PPV was 0.5 (95% CI, 0.03-0.98) in patients with clinical suspicion of IA. In conclusion, prevalence of IA in patients with liver cirrhosis seems to be low. Targeted GM testing in case of clinical suspicion of IA may be associated with markedly higher PPVs when compared to universal GM screening in patients with liver cirrhosis.

In the present study the spectrum and the incidence of fungi in potting soils and compost was investigated. Since soil is one of the most important biotopes for fungi, relatively high concentrations of fungal propagules are to be expected. For detection of fungi, samples of commercial soils, compost and soils from potted plants (both surface and sub-surface) were suspended and plated onto several mycological media. The resulting colonies were evaluated qualitatively and quantitatively. The results from the different sampling series vary, but concentrations on the surface of potted plants and in commercial soils are increased tenfold compared to compost and sub-surface soils. Median values range from 9.5 × 10(4) colony forming units (CFU)/g to 5.5 × 10(5) CFU/g. The spectrum of fungi also varies in the soils. However, all sampling series show high proportion of Aspergillus and Penicillium species, including potentially pathogenic species such as Aspergillus fumigatus. Cladosporium, a genus dominant in the ambient air, was found preferably in samples which were in contact with the air. The results show that potentially pathogenic fungi are present in soils. Immunocompromised individuals should avoid handling soils or potted plants in their immediate vicinity.

Objectives: Fulfilment of host factors defined by the revised European Organization for the Research and Treatment of Cancer/Mycoses Study Group (EORTC/MSG) criteria is required for establishing the diagnosis of possible or probable invasive fungal infection (IFI). This case‐control study evaluates EORTC/MSG host factors among patients with haematological malignancies. Methods: Fifty-eight patients with haematological malignancies who developed probable (n¼38) or proven (n¼20) IFI over a 5 year period were retrospectively evaluated regarding EORTC/MSG host factors. Results were compared with those obtained from patients with haematological malignancies who did not develop IFI (116 patients who received systemic antifungal prophylaxis or empirical therapy and 116 patients who did not; all data collected in 2010). Results: Fourteen patients had invasive yeast infection and 44 patients had invasive mould infection (IMI). Prolonged neutropenia (35/58, 60% versus 29/116, 25%), prolonged systemic corticosteroid (cut-off 21 days: 13/58, 22% versus 6/116, 5%; cut-off 14 days: 18/58, 31% versus 9/116, 8%) and T cell suppressive therapy (35/44, 80% versus 69/116, 59%) were significantly associated with development of IFI/IMI in our cohort. Previous allogeneic stem cell transplantation (SCT; .6 months prior to episode) was not significantly associated with development of IMI (8/44, 18% versus 22/116, 19%), while recent SCT (,6 months prior to episode) was (11/44, 25% versus 12/116, 10%). Conclusions: We conclude that host factors according to revised EORTC/MSG criteria were significantly associated with the development of IFI/IMI in our patients. Previous allogeneic SCT was not a predisposing host factor for the development of IMI. Concerning prolonged corticosteroid treatment, a cut-off of 14 days seems preferable to the proposed cut-off.

Summary Phaeohyphomycosis is a distinct mycotic infection of the skin or internal organs caused by darkly pigmented (dematiaceous) fungi, which are widely distributed in the environment. Phaeohyphomycosis is most frequently an opportunistic infection in immunosuppressed patients (HIV, corticotherapy, transplant patients) or is frequently associated with chronic diseases and diabetes. The spectrum of the disease is broad and includes superficial infections, onychomycosis, subcutaneous infections, keratitis, allergic disease, pneumonia, brain abscesses and disseminated disease. Rarely, immunocompetent patients may be affected. We describe two new cases of subcutaneous phaeohyphomycosis in immunocompetent patients: in the first patient, the causative agent was Exophiala jeanselmei, a common cause of phaeohyphomycosis; and in the second, Cladophialophora carrionii, which could be identified by culture. Cladophialophora carrionii is mainly the aetiological agent of chromoblastomycosis and only rarely the cause of phaeohyphomycosis. The first patient was treated with surgical excision and oral itraconazole, and the second patient responded to oral itraconazole only. Lesions improved in both patients and no recurrence was observed at follow-up visits.

Summary Heat shock proteins (Hsp) are highly conserved molecules, which are both constitutively expressed and up-regulated in response to various stress conditions. In particular, fungal Hsp60 can act as immunodominant antigens and facilitate powerful immunological properties. A possible cellular heat shock response was investigated in eight fungi (Aspergillus fumigatus, Aspergillus terreus, Penicillium chrysogenum, Cladosporium cladosporioides, Scedosporium apiospermum, Trichophyton mentagrophytes, Candida albicans and Saccharomyces cerevisiae). Fully automated RNA extraction was followed by quantitative real-time RT-PCR targeting fungus-specific Hsp60 mRNA and sequencing of the amplicon. Levels of temperature-dependent gene expression were evaluated and rates of similarity and identity were compared. While Hsp60 mRNA was constitutively expressed in all the samples tested, a temperature-dependent induction was not shown in C. cladosporioides. In the 80-amino acid fragment from the hypothetical protein, 66% of the amino acids were identical, 20% showed a conserved and 8% a semi-conserved substitution. Our findings should contribute to a better understanding of host–pathogen relationship and suggest that fungal Hsp60 under temperature-related stress conditions might act as an immunogenic trigger in orchestrating fungi-related diseases.

Brain infections caused by fungi of the Pseudallescheria / Scedosporium-complex weeks or months after a near drowning event should be considered, since mortality rate is high and specific diagnostic methods are necessary for fungal detection. Voriconazole is the only authorised antimycotic drug available for the treatment of these infections in Germany, but other antimycotics may also be indicated for alternate or combined treatment strategies. Consultation of a specific reference laboratory is recommended in comparable relevant cases.

Candida biofilms on indwelling devices are an increasing problem in patients treated at intensive care units. The goal of this study was to examine the occurrence and frequency of these biofilms. A total of 172 catheters were collected from 105 male and 67 female patients (the age range of both patient groups was from 3 weeks to 98 years old). The catheters were incubated on blood agar plates and the resulting yeast colonies were subsequently identified. Furthermore, pieces of catheters were fixed, dried and sputter coated with gold for investigation with scanning electron microscopy (SEM). Yeasts were recovered from significantly more catheters obtained from men than from women (chi(2): n = 67; P0.01). In SEM, 56.4% catheters turned out to be positive for biofilm formation. Again catheters from male patients were statistically significant (chi(2): n = 40; P0.01) more often positive than those from women. Candida albicans (71.1%) was the most common species isolated from the catheters, followed by C. glabrata (10.3%), C. parapsilosis (8.2%) and C. tropicalis (5.2%). Based on the results of this investigation, the epidemiology of Candida biofilms on indwelling devices seems to be a promising target for future investigations.

The genus Roesleria was introduced with the single species Roesleria hypogaea (current name: R. subterranea) by Thümen in 1877. The species was originally described from roots of grapevine and recognised as a facultative root-rotting parasite. The mazaediate ascoma with evanescent asci led to the assumption that Roesleria would be an ally of mazaediate lichens. In this study we calculate 28S, 18S as well ITS1-5.8S-ITS2 rDNA phylogenies. These data indicate that Roesleria is closely related to Hymenoscyphus (Helotiales). In contrast to other members of the Helotiales, the ascospores of Roesleria are passively released, and a distinct apical apparatus cannot be observed by LM. Electron optic investigations have also not elucidated whether the asci of Roesleria are equipped with a rudimentary apical apparatus or not. The passive release of the ascospores is discussed as an apomorphic character that evolved as adaptation to a hypogeous living.

Fungi, above all filamentous fungi, can occur almost everywhere, even in water. They can grow in such a quantity in water that they can affect the health of the population or have negative effects on food production. There are several reports of fungal growth in water from different countries, but to our knowledge none from Austria so far. The aim of this study was to gain an overview of the spectrum of filamentous fungi and yeasts in drinking water systems. Thirty-eight water samples from drinking water and groundwater were analysed. Fungi were isolated by using membrane filtration and plating method with subsequent cultivation on agar plates. The different taxa of fungi were identified using routine techniques as well as molecular methods. Fungi were isolated in all water samples examined. The mean value for drinking water was 9.1 CFU per 100 ml and for groundwater 5400 CFU per 100 ml. Altogether 32 different taxa of fungi were found. The taxa which occurred most frequently were Cladosporium spp., Basidiomycetes and Penicillium spp. (74.6%, 56.4% and 48.7%, respectively). This study shows that drinking water can be a reservoir for fungi, among them opportunists, which can cause infections in immunosuppressed patients.

Heat shock proteins or chaperones are found in mitochondrial and cytosolic compartments of cells. They are responsible for the correct folding of proteins and are up-regulated in reaction to various stressors. In addition, when released or presented on the surface of cells, they may play an important role in inflammatory and immunomodulating processes. To identify and characterize hsp60 in the common environmental mold Alternaria alternata, the fungus was cultivated and incubated at different temperatures to induce a possible heat shock response. Fully automated RNA extraction was followed by quantitative real-time RT-PCR targeting A. alternata specific Hsp60 mRNA and subsequent sequencing of the amplicon. While Hsp60 mRNA was constitutively expressed in all samples tested, a temperature-dependent expression of Hsp60 mRNA was observed. Sequencing revealed an identity of more than 85% to other fungal hsp60, indicating the existence of this protein in A. alternata.

Molds are found almost everywhere in the environment. Their airborne propagules (conidia, spores, hyphal fragments) can--under certain circumstances--cause a variety of health problems like mycotic infections, allergies, asthma, irritations or toxic syndromes.

Fungal spores are transported across great distances in the outdoor air and are also regularly found indoors. Building conditions and behavior-related problems in apartments may lead to massive growth of mold within a very short period of time. The aim of this study was to evaluate whether the visible growth of mold indoors influences the concentration of fungal spores in the air as well as the variety of their species. Samples were collected from 66 households in Austria. For each sampling, the corresponding outdoor air was measured as reference value. The size of the visible mold growth was categorized in order to correlate the extent of mold growth with the concentration of airborne spores as well as the fungal genera. In order to determine fungal spore concentrations in the air, the one-stage MAS-100® air sampler was used. Malt extract agar (MEA) and dichloran glycerol agar (DG18) plates were used as culture media. The total colony forming units (CFU) per m3 were determined. The fungi were identified from the isolated colonies. The results show that in apartments visibly affected by mold, the median values were significantly higher than those of apartments without visible mold growth. The extent of visible mold growth is significantly correlated with both concentration of fungal spores (p

Eosinophilic granulocytes (Eos) are found in great numbers both in the tissue and in the mucus of patients suffering from chronic rhinosinusitis with polyposis (ECRS). Interleukin-16 (IL-16) is known as a highly potent chemotactic and chemoattractant molecule (ED 10-11) for Eos. In an open, explorative, controlled study we examined the presence of IL-16 in mucosa tissue, mucus and serum in patients suffering from ECRS and its association to Eos activation. Tissue and nasal mucus specimen from 10 previously untreated, non allergic ECRS-patients undergoing paranasal sinus surgery and from 10 healthy non sinusitis subjects, undergoing nasal surgery because of anatomic nasal obstruction were investigated by real-time (RT-) PCR targeting human IL-16 mRNA. Haematoxylin-eosin (HE) staining and immunohistochemistry of formalin embedded tissue and mucus were applied for detection and determination of the proportion of activated Eos (aEos) and IL-16. Serum IL-16 was analyzed by enzyme-linked-immunosorbent assay (ELISA). IL-16 mRNA and IL-16 protein levels were elevated in nasal mucus, polyp tissue and in the serum of ECRS patients compared to healthy controls. There was a high proportion of aEos in ECRS patients compared to healthy subjects. Serum IL-16, IL-16 mRNA expression and IL-16 protein in mucus and tissue specimens were significantly associated with the presence of aEos in polyps of ECRS patients. Immunohistochemically IL-16 protein was mainly expressed in aEos, mast cells, lymphocytes and epithelial cells. In conclusion our data indicate that IL-16 may stimulate the migration and persistence of activated Eos in ECRS. IL-16 production in ECRS patients is not mediated by Immunglobuline-E (IgE).

Background Requirements on tissue fixatives are getting more demanding as molecular analysis becomes increasingly relevant for routine diagnostics. Buffered formaldehyde in pathology laboratories for tissue fixation is known to cause chemical modifications of biomolecules which affect molecular testing. A novel non-crosslinking tissue preservation technology, PAXgene Tissue (PAXgene), was developed to preserve the integrity of nucleic acids in a comparable way to cryopreservation and also to preserve morphological features comparable to those of formalin fixed samples. Methods Because of the excellent preservation of biomolecules by PAXgene we investigated its pathogen inactivation ability and biosafety in comparison to formalin by in-vitro testing of bacteria, human relevant fungi and human cytomegalovirus (CMV). Guidelines for testing disinfectants served as reference for inactivation assays. Furthermore, we tested the properties of PAXgene for detection of pathogens by PCR based assays. Results All microorganisms tested were similarly inactivated by PAXgene and formalin except Clostridium sporogenes, which remained viable in seven out of ten assays after PAXgene treatment and in three out of ten assays after formalin fixation. The findings suggest that similar biosafety measures can be applied for PAXgene and formalin fixed samples. Detection of pathogens in PCR-based diagnostics using two CMV assays resulted in a reduction of four to ten quantification cycles of PAXgene treated samples which is a remarkable increase of sensitivity. Conclusion PAXgene fixation might be superior to formalin fixation when molecular diagnostics and highly sensitive detection of pathogens is required in parallel to morphology assessment.

Vulvovaginal candidasis (VVC) is a common disease. The majority of cases is caused by Candida albicans, but in recent years an increase has been observed in the frequency of non-albicans Candida infections, especially due to C. glabrata and C. tropicalis. The aim of the study was to assess the prevalence of non-albicans Candida infections in patients suffering from VVC. Therefore, the statistical data of culture-confirmed VVC ascertained at the Institute of Hygiene (Medical University Graz) have been studied. Altogether, 10,463 samples from patients with vulvovaginal complaints were analysed in the years 2000-2004, a number of 3184 proved to be culture-positive for yeast. Candida albicans was the most prevalent cause in 87.9% of all cases. Non-albicans Candida yeast were detected in 12.1%, mainly C. glabrata and Saccharomyces cerevisiae. During a 1-year period 185 patients showed more than one episode of VVC. Patients aged 21-40 years were significantly more prone to suffer from VVC compared with other age-related groups.

Background Fungal spores are frequent in air and their occurrence in the nasal mucus appears to be a common finding within the adult population, as we were able to show in recent studies. 91,3 % of CRS patients but also healthy controls grew positive fungal cultures out of their nasal mucus. The potential role of fungal elements in nasal mucus for the pathogenesis of CRS, with or without polyposis, is currently investigated intensely and discussed very controversially. However, it was still unknown, as of when fungi could be cultured from nasal mucus in humans. We attempted to identify this point of time, in the nasal mucus of neonates. Methods In our study we examined nasal mucus from 30 neonates immediately after birth, on the first and fourth day post partum, and after two and four months of life. The samples obtained with sterile cotton swabs were cultured on agar plates. Fungal cultures were identified either conventionally by microscopy or with molecular techniques. To show whether fungi in nasal mucus of newborns were acquired by contamination during birth, mucus of the maternal vagina was examined as well. Results Just after birth we found in 6 of 30 (20 %) of our neonates positive fungal cultures out of their nasal mucus, in 3 of them Candida albicans, probably due to contamination passing the maternal vagina as cultures of vaginal mucus of their mothers were positive for Candida albicans too. Positive fungal cultures were obtained in 2 of 29 (7 %) neonates on the second and in 4 of 26 (15 %) neonates on the fifth day of life. In all our cases initial presence in nasal mucus contamination just after birth or on the second day of life was limited to one day only. None of the 12 of 30 (40 %) neonates with positive fungal cultures from nasal mucus in the first 5 days of life showed clinical symptoms of nasal fungal colonisation. Besides Candida albicans, Penicillium sp., Cladosporium cladosporioides, Acremonium polychromum, Beauveria bassiana and Epicoccum nigrum could be detected in the first 5 days of life. After the second month of life, examination of nasal mucus yielded positive fungal cultures in 8 of 11 (72 %), after four months even 17 of 18 (94 %) of babies, with a wide array of different species. Conclusions Fungi can be cultured from nasal mucus as soon as contact with the environmental air exists. Furthermore, a transfer of fungi from the mother's birth canal into the nose during birth is possible. Presence of fungal spores is common but not persistent in the nose of babies in the first days of life. However, after four months the situation is similar to the one in adults: fungal cultures can be obtained from almost everyone's nose. Therefore fungal spores must be considered a normal content of nasal mucus. Fungal spores are inhaled with every breath, some stick to the mucus, are transported to the nasopharynx and swallowed. This does not cause any clinical symptoms and is therefore not a pathological finding at all.

BACKGROUND Chronic Rhinosinusitis (CRS) is the most common chronic disease in the United States. Though for Europe no data are available, we have to assume that the situation is similar. Although the disease is defined very well by clinical symptoms, up to date the etiology and pathogenesis of chronic rhinosinusitis are unknown. CRS is considered to be multifactorial, with thickening of the mucosa and formation of polyps as an end stage of the disease. Treatment of choice includes corticosteroids and/or endoscopic or microscopic surgery. Antibiotics only help, if there is an acute bacterial exacerbation of the disease. They are not able to cure chronic rhinosinusitis per se. In 1999 Mayo Clinic researchers published data concerning the incidence of so-called "allergic" fungal sinusitis (AFS) in their patients suffering from chronic rhinosinusitis, demonstrating the majority of patients investigated presenting those criteria. Our own initial data from 2000 confirmed their findings. MATERIAL AND METHODS In an open prospective study fungal cultures were obtained from nasal mucus of 238 consecutive patients suffering from CRS. As control group acted 23 members of our staff, who did not show any evidence for CRS. In addition, in 37 CRS patients surgical specimens (mucus and tissue) were investigated histologically for evidence of eosinophilic granulocytes and fungal elements. RESULTS Using new techniques for fungal detection in culture and histology as proposed by Mayo Clinic researchers, positive detection of fungal cultures of the mucus of our CRS patients developed from 7 % in the past up to 87 % at present. 91.3 % of our control group yielded a positive result in fungal culture. Histologically, eosinophilic clusters were evident in 94.6 % and fungal elements were detected in 75.5 % within the mucus of 37 surgical CRS patients. Overall, 89.2 % of our surgical patients thus fulfilled the criteria of so-called AFS. Compared to our findings in the past, our latest results show an increase of 80 % in detection of fungal elements in our CRS patients. In all we were able to identify 654 positive fungal cultures in 238 CRS patients and 23 healthy controls respectively. 88 different genera grew, with 2.4 different species per patient and 3.1 different species per healthy control, on average. CONCLUSION Utilizing new techniques of fungal culturing out of the mucus of CRS patients and healthy controls, the number of positive fungal cultures increased dramatically from 7 % to 87 % in our patients and 91.3 % in healthy controls respectively. To obtain these results it is crucial to perform special techniques of mucus sampling and pretreatment for culturing as well as for histological investigations. Our results show, that with suitable techniques fungi can be identified in almost everybody's nose, CRS patient or healthy. When inhaled, those airborne fungi are only "in transit" through the nose. Positive fungal cultures from nasal secretion therefore have to be considered normal findings. The reason for this delayed recognition has to be attributed to our inadequate methods in the past. In contrast to healthy controls, clusters of eosinophils and fungal elements are present simultaneously within the mucus of CRS patients and appear to be a marker of the disease.

RationaleInvasive pulmonary aspergillosis has been increasingly reported in nonneutropenic patients, including those with underlying respiratory diseases.ObjectivesWe compared the diagnostic performances of galactomannan, 1,3-β-D-glucan, and Aspergillus-specific lateral-flow device tests with that of conventional culture by using bronchoalveolar lavage fluid samples from patients with underlying respiratory diseases.MethodsWe analyzed 268 bronchoalveolar lavage samples from 221 patients with underlying respiratory diseases (and without hematologic malignancy or previous solid organ transplantation) that were collected for routine microbiological workup between February 2012 and May 2014 at the University Hospital of Graz, Austria. Invasive pulmonary aspergillosis was defined according to European Organization of Research and Treatment of Cancer/Mycoses Study Group criteria modified for patients with respiratory diseases.Measurements and main resultsThirty-one patients (14%) had probable or proven, 25 possible, and the remaining 165 patients no invasive pulmonary aspergillosis. Probable/proven aspergillosis was associated with a significantly higher (P = 0.034) 30-day mortality rate of 32%. Sensitivities, specificities, and diagnostic odd ratios differed markedly between galactomannan (cut-off 0.5: optical density index, 0.97, 0.81, 124.4; cut-off 1.0: 0.97, 0.93, 422.1; cut-off 3.0: 0.61, 0.99, 109.8), β-D-glucan (cut-off 80 pg/ml: 0.90, 0.42, 6.57; cut-off 200 pg/ml: 0.70, 0.61, 3.7), lateral-flow device tests (0.77, 0.92, 41.8), and mycological culture (0.29, 0.97, 14).ConclusionsProbable or proven invasive pulmonary aspergillosis was diagnosed in 14% of our study population and associated with significantly higher 30-day mortality rates. Although the performance of β-D-glucan was limited by low specificity and that of mycological culture by low sensitivity, the Aspergillus lateral-flow device seems to be a promising alternative to galactomannan testing, which remains the diagnostic gold standard for aspergillosis. Clinical trial registered with www.clinicaltrials.gov (NCT 02058316).

BACKGROUND: The interplay between Candida species and pattern recognition receptors, interleukins, kynurenine, and T cells has been studied in murine and ex vivo human studies, but data are lacking from patients with invasive fungal infections. Interleukin 17A (IL-17A) is considered an important component in host defense against Candida infections and is modulated by Candida-induced impairment of tryptophan-kynurenine metabolism. METHODS: Dectin-1, Toll-like receptor 2, and Toll-like receptor 4 expression; regulatory T cell (Treg) percentages; and interleukin 6, interleukin 10, IL-17A, interleukin 22, interleukin 23, interferon γ, kynurenine, and tryptophan levels were determined in candidemic patients and compared to levels in noncandidemic patients who are in the intensive care unit (ICU) and receiving antibiotic therapy and those in healthy controls, both with and without Candida colonization. RESULTS: Candidemic patients had significantly higher IL-17A and kynurenine levels, compared with noncandidemic patients, including Candida-colonized ICU patients and healthy controls. Within candidemic patients, time-dependent elevation of IL-17A and kynurenine levels was detected. IL-17A areas under the curve for differentiation between patients with early candidemia and those without candidemia (ICU patients, including Candida-colonized patients, and healthy controls) were between 0.94 (95% confidence interval [CI], .89-.99) and 0.99 (95% CI, .99-1). CONCLUSIONS: Candidemic patients had significantly higher IL-17A and kynurenine levels, compared with noncandidemic patients. The statistically significant association between IL-17A and kynurenine levels and candidemia suggests their potential as biomarkers for anticipation of invasive candidiasis. CLINICAL TRIALS REGISTRATION: NCT00786903.

Testing for serum galactomannan (GM) has been established as an important method for diagnosing invasive aspergillosis (IA); however, limited data exist regarding the application of urine GM testing. The objective of this study was to evaluate the performance of GM screening of urine specimens and to compare results with serum GM. The study was performed between July 2012 and March 2013 in adult patients with underlying hematological malignancies who were hospitalized at the Medical University of Graz, Austria. Serum and urine screening samples were collected and tested twice weekly (always on the same day). In total, 242 serum samples and a similar number of urine samples were collected from 75 patients. A total of 21/242 (8.7%) serum samples from 13 patients were GM positive. Sensitivity, specificity, positive predictive value, and negative predictive value using a 0.1 optical density index cutoff for urine samples (compared with same-day serum results) were as follows: 47.6%, 86%, 24.4%, and 94.5%, respectively. In 8/10 patients with probable IA, at least one positive GM result was found with this cutoff. After calculating clinical performance of the urine GM test, we found that sensitivity increased to 71.4% and specificity to 88.2%. Spearman-Rho correlation analysis revealed a significant positive correlation between serum and urine samples (P 0.001; ρ = 0.252). In conclusion, GM detection in urine might be a promising method for IA screening. However, further studies are needed.

Galactomannan detection in bronchoalveolar lavage (BAL) fluid samples (GM test) is currently considered the gold standard test for diagnosing invasive pulmonary aspergillosis (IPA). The limitations, however, are the various turnaround times and availability of testing. We compared the performance of GM testing with that of conventional culture, an Aspergillus lateral-flow-device (LFD) test, a beta- d -glucan (BDG) test, and an Aspergillus PCR assay by using BAL fluid samples from immunocompromised patients. A total of 78 BAL fluid samples from 78 patients at risk for IPA (74 samples from Graz and 4 from Mannheim) collected between December 2012 and May 2013 at two university hospitals in Austria and Germany were included. Three patients had proven IPA, 14 probable IPA, and 17 possible IPA, and 44 patients had no IPA. The diagnostic accuracies of the different methods for probable/proven IPA were evaluated. The diagnostic odds ratios were the highest for the GM, PCR, and LFD tests. The sensitivities for the four methods (except culture) were between 70 and 88%. The combination of the GM (cutoff optical density index [ODI], >1.0) and LFD tests increased the sensitivity to 94%, while the combination of the GM test (>1.0) and PCR resulted in 100% sensitivity (specificity for probable/proven IPA, 95 to 98%). The performance of conventional culture was limited by low sensitivity, while that of the BDG test was limited by low specificity. We evaluated established and novel diagnostic methods for IPA and found that the Aspergillus PCR, LFD, and GM tests were the most useful methods for diagnosing the disease by using BAL fluid samples. In particular, the combination of the GM test and PCR or, if PCR is not available, the LFD test, allows for sensitive and specific diagnosis of IPA.

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Zusammenfassung. 27 Trichophyton verrucosum-Stamme aus 14 Rinderbestanden in Thuringen und Mecklenburg-Vorpommern wurden unter Einbeziehung von sechs T. verrucosum-Referenzstammen, unter anderem auch die sogenannten album-, discoides- und ochraceum-Varietaten, kulturmorphologisch und molekularbiologisch (PCR-Fingerprinting, AFLP-Analyse sowie Sequenzieung der ITS-Region der rDNA) gepruft. Trotz groser kulturmorphologischer Variabilitat der lsolate und ihrer moglichen Zuordnung zu vier verschiedenen Kolonietypen waren alle gepruften T. verrucosum-Stamme genotypisch identisch. Auch die zwei mit Gelbpigment wachsenden Feldisolate, die moghcherweise als ochraceum-Varietat angesehen werden konnten, sowie der ochrceum-Referenzstamm konnten mittels der drei genannten molekularbiologischen Methoden nicht differenziert werden. Somit ergeben sich weder Hinweise fur die eigenstandige taxonomische Stellung der drei T. verrucosum-Varietaten noch konnte die vermutete Infektion mit ochraceum-Stammen in Rinderbestanden als Ursache fur das Versagen der Impfprophylaxe mit verschiedenen T. verrucosum-Impfstoffen und fur das gehaufte Auftreten sogenannter Impfversager bestatigt werden. Es wird vielmehr vermutet, das neben der unzureichenden Impfstammpflege vor allem Anwendungsfehler, insbesondere in der luckenlosen, systematischen Immunprophylaxe in der Rinderherde, ursachlich fur das gehaufte Auftreten erkrankter Rinder in gegen Trichophytie geimpften Bestanden verantwortlich sind. Summary. Twenty-seven strains of Trichophyton verrucosum from 14 cattle herds in the Federal States of Thuringia and Mecklenburg-Vorpommern were examined by culture morphological and molecular biological (PCR fingerprinting, AFLP analysis sequencing of ITS region) methods. Six reference strains of the same species, among them the so-called album andochraceum varieties, were also included. Despite great variability in terms of culture morphology, which suggested their possible classification into 4 different colony types, all T. verrucosum isolates were genotypically almost identical. Even the 2 field isolates growing with yellow pigment, which could possibly be regarded as belonging to the ochraceum variety, could not be differentiated using molecular biological methods. The results do not provide indications of a separate taxonomic position of the 3 T. verrucosum varieties. Furthermore, there is no evidence confirming the suspected infection of cattle herds with ochraceum strains as the cause of the failure of immune prophylaxis using various T. verrucosum vaccines. The frequent occurrence of animals not responding to vaccination could not be explained either. It should be assumed that the main factors responsible for this situation include poor handling of the vaccine strains and errors in application, especially the absence of continuous and systematic immune prophylaxis in the herds.