Tissue characterization of the medical fungus Hericium coralloides by focus-variation microscopy.

In this study, the potential of focus-variation microscopic imaging was evaluated in a study of morphological patterns of the potential medicinal fungus Hericium coralloides (Basidiomycota). We created three-dimensional reconstructions and visualizations using the imaging technique on a fresh H. coralloides basidioma. The aim was to approximate the spore dispersal efficiency of this basidiomata type regarding the investment of tissue biomass and its reproductive output (production of basidiospores). Results were correlated with published data gained from magnetic resonance imaging and micro-computed tomography. It is demonstrated that focus-variation microscopic imaging results in a more distinct picture of the morphology of the edible and potentially medicinal H. coralloides basidiomata. However, a direct measurement of spore production was not possible. Spore production could only be estimated in combination with a mathematical model because the surface was not directly measurable due to the cellular heterogeneity. However, focus-variation microscopic imaging allows a better and faster estimation of spore production compared with the published methods. Furthermore, it was found that a scanning resolution of 5× is sufficient for determining the fungal surface precisely because at a higher resolution artifacts occur, resulting in adulteration of the image.

Morphological and tissue characterization of the medicinal fungus Hericium coralloides by a structural and molecular imaging platform.

In this study the potential of new imaging techniques such as Magnetic Resonance Imaging (MRI), Matrix-Assisted Laser Desorption/Ionization (MALDI) profiling mass spectrometry ("MALDI Profiling") and Fourier Transform Infrared (FTIR) spectroscopic imaging was evaluated to study morphological and molecular patterns of the potential medicinal fungus Hericium coralloides. For interpretation, the MALDI profiling, FTIR imaging and MRI results were correlated with histological information gained from Scanning Electron Microscopy (SEM) and Light Microscopy (LM). Additionally we tested several evaluation processes and optimized the methodology for use of complex FTIR images to monitor molecular patterns. It is demonstrated that the combination of these spectroscopic methods enables to gain a more distinct picture concerning morphology and distribution of active ingredients. We were able to obtain high quality FTIR imaging and MALDI-profiling results and to distinguish different tissue types with their chemical ingredients. Beside this, we have created a 3-D reconstruction of a mature Hericium basidioma, based on the MRI dataset: analyses allowed, for the first time, a realistic approximation of the "evolutionary effectiveness" of this bizarrely formed basidioma type, concerning the investment of sterile tissue and its reproductive output (production of basidiospores).

Biodiversity and concentration of airborne fungi in a hospital environment.

The biodiversity and concentration of airborne fungi were monitored over a period of 6 months in a special-care unit of a hospital. Air sampling was performed in a corridor that was also accessible to visitors and in an adjacent bone-marrow transplantation (BMT) unit using an air sampler and two isolation media. Altogether, 98 fungal species could be identified, among them Aspergillus fumigatus and A. terreus as well as 48 other species reported as potential pathogens. The average contamination values of the corridor air ranged from 124 to 485 cfu m-3. Neither the degree of fungal air contamination nor the species composition inside the special care unit differed from those found in the corridor. By means of data obtained with a light-activated sensor, a possible influence of human activities on diurnal changes of fungal propagule concentration was shown.

The mycobiota of speck, a traditional Tyrolean smoked and cured ham.

Speck is a ham specialty product traditionally produced in South Tyrol (Italy) and North Tyrol (Austria) by farmers, butcheries, and meat industries. To date, nothing has been learned about fungi associated with this smoked and cured meat. Therefore, it was the main objective of this study to assess the typical mycobiota of Speck in relation to the different production types and the geographic provenance. A total of 121 Speck samples from North Tyrol and South Tyrol was analyzed. From 63 isolated fungal species, only a few can be regarded as typical colonizers: Eurotium rubrum and Penicillium solitum were the dominating species in all types and parts of Speck (crust, meat, and fat). Eight other Penicillium spp. were relatively frequent. The species diversity increased from industrially produced Speck to products from butcheries and farmers, and it was higher in all types of South Tyrolean products. Among the typical mycobiota, Penicillium verrucosum, Penicillium canescens, and Penicillium commune are known as potentially mycotoxigenic.

Identification of illudins in Omphalotus nidiformis and Omphalotus olivascens var. indigo by column liquid chromatography-atmospheric pressure chemical ionization tandem mass spectrometry.

Reversed-phase liquid chromatography was used to separate toxins in mushrooms of the genus Omphalotus. Crude ethyl acetate extracts of cultures were injected directly onto a 150 x 2 mm I.D. column packed with 3 microns octadecylsilica and eluted with a gradient of acetonitrile in 0.1% aqueous acetic acid at a flow-rate of 200 microliters/min. Monitoring of the column effluate by atmospheric pressure ionization tandem mass spectrometry allowed the identification of the toxins. The fungal toxins illudin M and illudin S were detected and identified for the first time in cultures of the Australian Omphalotus nidiformis and the North American Omphalotus olivascens var. indigo (Boletales, Basidiomycetes) and confirmed the valuable taxonomic character of illudins for the genus Omphalotus.

Orellanine poisoning: rapid detection of the fungal toxin in renal biopsy material.

BACKGROUND: Mushroom poisoning by some species of the Cortinarius (Agaricales) often lead to irreversible renal failure caused by the nephrotoxin orellanine. In 1994 and 1995, six poisoning outbreaks involving ten individuals in Northern Italy and in Austria were investigated. METHODS: A total of 87 clinical samples (urine and blood samples including renal biopsy material of three patients) were examined for the presence of orellanine by thin layer chromatography. RESULTS: Orellanine can be detected after a relatively long period following poisoning by performing a simple thin layer chromatography technique using small quantities of renal biopsy material. No toxin was found in urine or blood samples. CONCLUSIONS: Orellanine is rapidly concentrated in the kidneys in a relatively soluble form and cannot be detected in urine, blood and dialysis fluids at the time when first symptoms appear.