NOWE STANOWISKO SOPLÓWKI BUKOWEJ HERICIUM CORALLOIDES (Scop.) Pers. w WIELKOPOLSCE

Aim. This study was aimed to examine influence of сytokinin fraction of basidiomycete Hericium coralloides on the spontaneous and induced phagocytic activity of murine peritoneal macrophages. Materials and methods. Mononuclear phagocyte (macrophage) fraction of peritoneal exudate of C57BL/6 mice was used. Macrophages were incubated under standard conditions at 37 °C, 100% humidity and 5% CO2 for 4 hours. Phorbol 12-myristate 13-acetate (PMA) was added to part of the samples to activate oxidative metabolism. Hericium coralloides was added in two concentrations that were 5 and 10 times lower than the IC50, defined as an antiproliferative effect on colon cancer cells. Incubation with samples was carried out for 2 hours. Results. When adding PMA, Hericium coralloides (0.017 and 0.035 μg/ml) and in the combination of PMA with Hericium coralloides, activation of reactive oxygen species (ROS) in peritoneal macrophages was revealed by 1.37-1.7 times, compared to the spontaneous activity of phagocytes. Conclusions. Thus, the effect of the cytokinin e.tract of the basidium fungus Hericium coralloides was manifested by an increase in the phagocytic activity of peritoneal macrophages as one of the possible mechanisms of immunomodulatory action.

The potential of 3-D nondestructive imaging techniques such as micro-computed tomography (micro-CT) was evaluated to study morphological patterns of the potential medicinal fungus Hericium coralloides (Basidiomycota). Micro-CT results were correlated with histological information gained from scanning electron microscopy (SEM) and light microscopy (LM). It is demonstrated that the combination of these imaging methods results in a more distinct picture of the morphology of the edible and potentially medicinal Hericium coralloides basidiomata. In addition we have created 3-D reconstructions and visualizations based on micro-CT imagery from a randomly selected part of the upper region of a fresh H. coralloides basidioma: Analyses for the first time allowed an approximation of the evolutionary effectiveness of this bizarrely formed basidioma type in terms of the investment of tissue biomass and its reproductive output (production of basidiospores).

Nowadays the studies of the urban fungal diversity with highlighting mostly macrofungal species of conservation concern is a rapidly growing field of interest. The present paper reports the first finding of the wood-inhabiting fungus Hericium coralloides (Scop.) Pers., which has been considered rare and threatened in Europe and is included in the Red List of Bulgarian fungi with Near Threatened category, in the Sofia city park Borisova gradina. The previous findings of the same species in Plovdiv and in the Sofia park Vrana together with the records of another close and threatened species of the same genus, considered as Endangered in Bulgaria, namely H. erinaceus (Bull.) Pers., in another city park of Sofia – Zapaden park, strongly suggests that both tooth fungi tend to be members of the urban mycota in Bulgaria despite their apparent rarity. This serves to prove the need to investigate currently under-utilized opportunity that cities with their distinct species assemblages present for biodiversity conservation, when they are recognized as hotspots for threatened species.

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.

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).