Delimitation and characterisation of Talaromyces purpurogenus and related species.

Taxa of the Talaromyces purpurogenus complex were studied using a polyphasic approach. ITS barcodes were used to show relationships between species of the T. purpurogenus complex and other Talaromyces species. RPB1, RPB2, ß-tubulin and calmodulin sequences were used to delimit phylogenetic species in the complex. These data, combined with phenotypic characters, showed that the complex contains four species: T. purpurogenus, T. ruber comb. nov. and two new species T. amestolkiae sp. nov. and T. stollii sp. nov. The latter three species belong to the same clade and T. purpurogenus is located in a phylogenetic distant clade. The four species all share similar conidiophore morphologies, but can be distinguished by macromorphological characters. Talaromyces ruber has a very distinct colony texture on malt extract agar (MEA), produces bright yellow and red mycelium on yeast extract sucrose agar (YES) and does not produce acid on creatine sucrose agar (CREA). In contrast, T. amestolkiae and T. stollii produce acid on CREA. These two species can be differentiated by the slower growth rate of T. amestolkiae on CYA incubated at 36 °C. Furthermore, T. stollii produces soft synnemata-like structures in the centre of colonies on most media. Extrolite analysis confirms the distinction of four species in the T. purpurogenus complex. The red diffusing pigment in T. purpurogenus is a mixture of the azaphilone extrolites also found in Monascus species, including N-glutarylrubropunctamine and rubropunctatin. Talaromyces purpurogenus produced four different kinds of mycotoxins: rubratoxins, luteoskyrin, spiculisporic acid and rugulovasins and these mycotoxins were not detected in the other three species.

[Quality assurance in the diagnosis of indoor fungi]. / Ringversuch "Innenraumrelevante Schimmelpilze".

In co-ordination with the Umweltbundesamt Berlin, the Landesgesundheitsamt Baden-Wurttemberg (LGA) initiated external quality assurance in the diagnosis of indoor fungi in autumn 2001. Four of six fungal strains commonly found indoors have to be fully identified (on the genus and species level). There are two distributions per year; the six distributions hitherto carried out resulted in correct identification by 46-89% of laboratories (40-71 participants, total 148). It is clear from the results that repeat participants were more successful. In addition to the pure cultures sent out we offered actual samples (two air samples, one wood material, one sample of house dust, hitherto); 43- 69% of participating laboratories also took part in this facultative investigation of actual samples and 29-62% were successful. Results that differed considerably revealed problems while treating and evaluating actual samples. Therefore, activities in this field should be enhanced. In conclusion, external quality assurance in the diagnosis of indoor fungi is a useful management aid in the maintenance and improvement of laboratory performance.

Isolation of Neosartorya pseudofischeri from blood: first hint of pulmonary Aspergillosis.

We report a case of invasive pulmonary aspergillosis caused by Neosartorya pseudofischeri S. W. Peterson [anamorph Aspergillus thermomutatus (Paden) S. W. Peterson]. The diagnosis was initially based on a positive blood culture for a strain isolated from a neutropenic patient by means of a BACTEC 9050 blood culture system. The final diagnosis was established based on X-ray and computer tomography scan results as well as the detection of Aspergillus antigen in the patient's serum.

Analysis of Fusarium causing dermal toxicosis in marram grass planters.

In the European coastal dunes, marram grass (Ammophila arenaria) is planted in order to control sand erosion. In the years 1986 to 1991, workers on the Wadden islands in the Netherlands planting marram grass showed lesions of skin and mucous membranes, suggesting a toxic reaction. Fusarium culmorum dominated the mycoflora of those marram grass culms that were used for planting. This plant material had been cut and stored for more than one week in the open. The Fusarium toxin deoxynivalenol (DON) was detected in the suspect marram grass culms. Isolated F. culmorum strains were able to produce DON in vitro in liquid culture as well as in experimentally inoculated wheat heads. Pathogenicity tests, toxin test as well as RAPD analysis showed that the F. culmorum strains were not specialized for marram grass but may form part of the West-European F. culmorum population infecting cereals and grasses. Storage on old sand-dunes with plant debris may have led to the high occurrence of F. culmorum and contamination with DON. Marram grass culms should be obtained from young plantings on dunes on the seaward slopes and cut culms should not be stored.

Fungal propagules in house dust. I. Comparison of analytic methods and their value as estimators of potential exposure.

The presence of viable mold propagules in house dust was investigated by 10 different analytic methods, in order to determine to what extent different results are obtained when different analytic methods are used. Moreover, the value of this measurement as an estimator of the potential exposure to fungi in epidemiologic studies was assessed. Floor and mattress dust was sampled in 60 homes in The Netherlands during autumn 1990. For investigation of the variability in time, sampling was repeated in 20 homes after 6 weeks. Each analytic method is characterized by a unique combination of culture medium, suspension medium, and dilution step. The highest mean number of colony-forming units (CFU)/g dust was obtained by suspension of at least 100 mg dust in a peptone or sucrose solution in a ratio of 1:50 (w/w), followed by 10-fold dilution and plating on DG18 agar (geometric mean (GM) approximately 60,000 CFU/g dust). The lowest mean number of CFU/g dust was obtained by direct plating of 30 mg dust on V8 agar (GM approximately 5300 CFU/g dust). The mean coefficient of variation of duplicate analyses varied from 11%, for suspension in sucrose and plating on DG18 agar, to 27%, for suspension and dilution in sucrose in combination with V8 agar. The highest mean number of species isolated was obtained by direct plating of 30 mg dust on DG18 agar (mean number of species: 17). Suspension and dilution on DG18 or V8 agars yielded an average of approximately six species. In duplicate analyses, the mean percentage of agreement for the species isolated varied from approximately 35%, for suspension and dilution, to 60%, for direct plating.(ABSTRACT TRUNCATED AT 250 WORDS)

Fungal propagules in house dust. II. Relation with residential characteristics and respiratory symptoms.

As part of a case-control study on the relation between home dampness and respiratory symptoms of children, house-dust samples were collected from bedroom floors and mattresses in 60 homes in The Netherlands. The house-dust samples were analyzed for the presence of fungal propagules by plating 30 mg of dust directly onto DG18 agar. A checklist and questionnaire were used to obtain information on the home characteristics and occupant behavior that may have an effect on the presence of fungal propagules in house dust. The geometric mean (GM) numbers of colony-forming units (CFU)/g dust collected from the floors was 8990. The number of CFU/g dust was significantly higher in dust from carpeted floors than in dust from smooth floors (GM, respectively, 12,880 CFU/g dust and 3530 CFU/g dust). The GM number of CFU/g dust collected from mattresses was 6760. Overall, the mean numbers of CFU/g dust collected from floors and mattresses were higher in bedrooms where damp spots mold growth, or both were observed. However, these differences were not statistically significant. The relation between home characteristics and the number of CFU/g dust of the most frequently isolated mold species (n = 17), including Alternaria alternata, Cladosporium cladosporioides, Penicillium brevicompactum, and Scopulariopsis brevicaulis, was also investigated. Only the type of flooring had a significant and consistent effect on the number of CFU/g floor dust of the different mold species.(ABSTRACT TRUNCATED AT 250 WORDS)

Presence of viable mould propagules in indoor air in relation to house damp and outdoor air.

The presence of viable mould propagules in indoor air was investigated using the N6-Andersen sampler in combination with DG18-agar, in relation to house damp (characterized with a checklist) and in relation to the presence of moulds in outdoor air. The first part of the study was conducted in 46 houses in the autumn of 1987, the second part in 84 houses in May 1989. Further, in the second part, the results obtained with settlement plates (OPD) were compared with those obtained with the N6-Andersen sampler. The number of CFU/m3 in the indoor and outdoor air varied widely. A large variety of mould genera and species was isolated. Species of Cladosporium, Penicillium and Wallemia predominated. The variability in time was high and the reproducibility of the measurements in terms of CFU/m3 and of species isolated was only moderate. The low predictive value of these measurements limits their use in epidemiological studies of the relationship between exposure to moulds and respiratory symptoms. Overall, the geometric mean concentration was somewhat higher outdoors than indoors. However, the clear differences found between the number of CFU/m3 belonging to different mould species in in- and outdoor air show that the presence of viable mould propagules in indoor air is not simply a reflection of the presence of moulds in outdoor air. The presence of moulds in indoor air was only weakly related to house damp as characterized by the checklist. High, statistically significant correlations were found between the CFU yield obtained with the OPD and the CFU/m3 yield obtained with the N6-Andersen sampler.(ABSTRACT TRUNCATED AT 250 WORDS)

Enumeration and identification of airborne viable mould propagules in houses. A field comparison of selected techniques.

A number of techniques for the enumeration and identification of viable mould propagules in the indoor air of houses were evaluated in order to document to what extent different results are obtained when different methods are used. A comparison was made between the results obtained with five commercially available air sampling devices (Slit-to agar sampler, N6-Andersen sampler, Surface Air System sampler, Reuter Centrifugal Air sampler, Gelatine Filter sampler) and a non-volumetric sampler (the Open Petri Dish), in combination with four culture media (malt extract agar, dichloran glycerol-18 agar, oxytetracycline glucose yeast extract agar and dichloran rose bengal chloramphenicol agar). The coefficients of variation were high (generally greater than 20%) for all combinations. Statistical analysis showed that the Slit sampler and the N6-Andersen sampler in combination with DG18 and MEA gave the best precision and the highest yield in terms of colony forming units per square cubic meter of air (CFU/m3) and number of species isolated.