The “Nietoperek” bat reserve located in Western Poland is one of the largest bat hibernation sites in the European Union with nearly 38,000 bats from 12 species. Nietoperek is part of a built underground fortification system from WWII. The aims of the study were (1) to determine the fungal species composition and changes during hibernation season in relation to bat number and microclimatic conditions and (2) evaluate the potential threat of fungi for bat assemblages and humans visiting the complex. Airborne fungi were collected in the beginning, middle and end of hibernation period (9 November 2013 and 17 January and 15 March 2014) in 12 study sites, one outside and 11 inside the complex. Ambient temperature (T _a) and relative humidity (RH) were measured by the use of data loggers, and species composition of bats was recorded from the study sites. The collision method (Air Ideal 3P) sampler was used to detect 34 species of airborne fungi including Pseudogymnoascus destructans (Pd). The density of airborne fungi isolated from the outdoor air samples varied from 102 to 242 CFU/1 m³ of air and from 12 to 1198 CFU in the underground air samples. There was a positive relationship between number of bats and the concentration of fungi. The concentration of airborne fungi increased with the increase of bats number. Analysis of other possible ways of spore transport to the underground indicated that the number of bats was the primary factor determining the number of fungal spores in that hibernation site. Microclimatic conditions where Pd was found (median 8.7 °C, min-max 6.1–9.9 °C and 100 %, min-max 77.5–100.0 %) were preferred by hibernating Myotis myotis and Myotis daubentonii; therefore, these species are most probably especially prone to infection by this fungi species. The spores of fungi found in the underground can be pathogenic for humans and animals, especially for immunocompromised persons, even though their concentrations did not exceed limits and norms established as dangerous for human health. In addition, we showed for the first time that the air in bats hibernation sites can be a reservoir of Pd. Therefore, further study in other underground environments and wintering bats is necessary to find out more about the potential threat of airborne fungi to bats and public health.

Osówka (Germ. Säuferhöhen), is one of the elements of the unfinished Nazi military complex called “Riese”. The total length of corridors of Osówka complex is about 1,700 m and its capacity amounts to 30,000 m3. As described geologically, Osówka is situated within the Sowie Mts. Massif which consists mostly of various gneisses with different structural characteristics, but with a constant mineral composition. The rock-forming minerals are feldspar (oligoclase), quartz, biotite and light micas, accesory minerals are garnet, sillimanite and dysten. Fine-grained shallow-sea deposits were probably a protolith of these rocks. The study aimed at first mycological evaluation of the air and the rocks in Osówka adit. The air samples were taken from one location outside the adit and from four locations inside of it. Mycological evaluation of the rocks inside the adit was performed using three different methods. Fifteen taxa of filamentous fungi were isolated from the internal air sampled, and several taxa - from the outside of the adit, whereas only eleven species were isolated from the rocks. Cladosporium spp. were the fungi most frequently isolated from internal atmosphere of the underground Osówka complex, and from the external air. On the other hand, the fungi most frequently isolated from the rocks were Aspergillus niger group (when using swab sampling procedure) and Mucor spp. (from debris and rinse sampling procedure).

Biscogniauxia nummularia (Bull.) Kuntze is a fungus which induces strip-cankers on beech, commonly referred to as charcoal canker. The symptoms of infection are visible on the host tree’s bark as elongated, blackish bark lesions on the trunk and branches. Recent years have shown that, due to climate change causing local epidemics, the species is increasing its economic impact in Mediterranean regions. Until recently, B. nummularia was considered rare and uncommon in central Europe. However, in the last few years it has been noticed more often, mostly in coniferous trees, which are out of B. nummularia’s host range. A similar situation has been observed with the closely related species Biscogniauxia mediterranea (De Not.) Kuntze, which prior to 2017 had not been observed in central Europe at all. This study shows the genetic diversity of mid-European strains of Biscogniauxia spp. (based on the ITS, TEF1, TUB2 and ACT regions) and, as the first in Europe, presents a molecular investigation of this species isolated from coniferous trees. It is also the first attempt at estimating the potential impact of this pathogenic fungus on European forestry management in the close future.

The internal transcribed spacer (ITS) region is regarded as a formal fungal primary barcode with a high probability of the correct identification for a broad group of fungi. ITS sequences have been widely used to determine many fungal species and analysis of rDNA ITS is still one of the most popular tools used in mycology. However, this region is not equally variable in all groups of fungi; therefore, identification may be problematic and result in ambiguous data, especially in some species-rich genera of Ascomycota. For these reasons, identification based on rDNA ITS is usually complemented by morphological observations and analysis of additional genes. Reliable species identification of Ascomycota members is essential in diagnosing plant diseases, verifying air quality and the effectiveness of agronomic practices, or analyzing relationships between microorganisms. Therefore, the present study aimed to verify, using specific examples, the extent to which ITS sequence analysis is useful in species identification of pathogens and saprobionts from Ascomycota and demonstrate problems related to such identification in practice. We analyzed 105 ITS sequences of isolates originating from air and plant material. Basic local alignment search tool (BLASTn) significantly contributed to the reliable species identification of nearly 80% of isolates such as Arthrinium arundinis, Beauveria bassiana, Boeremia exigua, Cladosporium cladosporioides, Epicoccum nigrum, Nigrospora oryzae, Sclerotinia sclerotiorum, or Sordaria fimicola and members of the genera Alternaria and Trichoderma. However, for most isolates, additional morphological observations, information regarding the isolate origin and, where possible, a PCR with species-specific primers were helpful and complementary. Using our practical approach, we determined that ITS-based species identification and comparative analysis with GenBank sequences significantly helps identifying Ascomycota members. However, in many cases, this should be regarded as suggestive of a taxon because the data usually require the use of additional tools to verify the results of such analysis.