Figure - uploaded by Rafał Ogórek
Content may be subject to copyright.
BLAST analysis of nucleotide sequences of PCR fungal products obtained in amplification reaction with ITS1 and ITS4 primers. All E values were 0.0.
Source publication
Underground ecosystems are one of the most inhospitable places for microorganism development and function. Therefore, any organic matter located in these areas can stimulate fungal growth. The main purpose of this study was to find the best solution to effectively preserve (without relapses) paleolithic bones of cave bear (Ursus spelaeus) exhibited...
Contexts in source publication
Context 1
... was expected, direct examination of the powdered bones (prepared using drill) did not show any fragments of hyphae (data not shown). We isolated a total of 11 fungal species from the bones ( Table 1). Macromorphology of colonies growing on different media (Figure 3) and micromorphology of fungal structures documented during the microscopic observations of LPCB stained preparations ( Figure 4) were consistent with what we observed during the direct microscopical examination of the bone scrapings, both presence of coenocytic as regularly septated hyphae. ...Context 2
... the photos taken after two weeks incubation of bones at 20 ± 1 • C under laboratory conditions (specified in Section 2). We isolated a total of 11 fungal species from the bones ( Table 1). Macromorphology of colonies growing on different media (Figure 3) and micromorphology of fungal structures documented during the microscopic observations of LPCB stained preparations ( Figure 4) were consistent with what we observed during the direct microscopical examination of the bone scrapings, both presence of coenocytic as regularly septated hyphae. ...Context 3
... four of eleven isolates, coenocytic hyphae and the fastest growth rate were observed. Together with microscopic observations, this allows us to classify We isolated a total of 11 fungal species from the bones ( Table 1). Macromorphology of colonies growing on different media (Figure 3) and micromorphology of fungal structures documented during the microscopic observations of LPCB stained preparations ( Figure 4) were consistent with what we observed during the direct microscopical examination of the bone scrapings, both presence of coenocytic as regularly septated hyphae. ...Context 4
... the isolates which were identified based on traditional, phenotypic methods were confirmed by molecular methods. The size of obtained PCR products were in the range of 504-648 bp (Table 1). Results of molecular identification were completely consistent with those obtained by traditional methods. ...Context 5
... of molecular identification were completely consistent with those obtained by traditional methods. All the nucleotide sequences obtained with primers ITS1 and ITS4 together with the corresponding sequences to which these showed the closest match are deposited in Genbank under accession numbers in Table 1. In all of the BLAST analysis done the E values were zero (0.0) and percent of query cover and identity were in the range of 96.0-100% and 94.71-100%, respectively. ...Citations
... Identification was performed by morphological and molecular methods [42][43][44][45][46][47][48][49][50][51][52][53][54][55]. In detail, a molecular analysis was performed, for which DNA was extracted from a 28-dayold culture on PDA using Bead-Beat Micro AX Gravity kit (A&A Biotechnology, Gdańsk, Poland) according to the manufacturer's instructions. ...
Despite speleomycological research going back to the 1960s, the biodiversity of many specific groups of micromycetes in underground sites still remains unknown, including keratinolytic and keratinophilic fungi. These fungi are a frequent cause of infections in humans and animals. Since subterranean ecosystems are inhabited by various animals and are a great tourist attraction, the goal of our research was to provide the first report of keratinophilic and keratinolytic fungal species isolated from three caves in Tatra Mts., Slovakia (Brestovská, Demanovská Lǎdová and Demanovská Slobody). Speleomycological investigation was carried out inside and outside the explored caves by combining culture-based techniques with genetic and phenotypic identifications. A total of 67 fungal isolates were isolated from 24 samples of soil and sediment using Van-breuseghem hair bait and identified as 18 different fungal species. The study sites located inside the studied caves displayed much more fungal species (17 species) than outside the underground (3 species), and the highest values of the Shannon diversity index of keratinophilic and keratinolytic fungi were noted for the study sites inside the Demänovská Slobody Cave. Overall, Arthroderma quadrifidum was the most common fungal species in all soil and/or sediment samples. To the best of our knowledge, our research has allowed for the first detection of fungal species such as Arthroderma eboreum, Arthroderma insingulare, Chrysosporium europae, Chrysosporium siglerae, Keratinophyton wagneri, and Penicillium charlesii in underground sites. We also showed that the temperature of soil and sediments was negatively correlated with the number of isolated keratinophilic and keratinolytic fungal species in the investigated caves.
... Firstly, the obtained fungi were identified using classic phenotypic methods according to available monographs and articles [33][34][35][36][37][38][39][40][41][42][43][44][45][46]. In the next step, a molecular analysis of the obtained fungal cultures was performed. ...
Most underground ecosystems are heterotrophic, fungi in these objects are dispersed in the air in the form of spores, and they may be potentially hazardous to mammals. Research in underground sites has focused on mesophilic airborne fungi and only a few concerned cold-adapted species. Therefore, the goal of our research was the first report of psychrophilic and psychrotolerant aeromycota in the Brestovská Cave using culture-based techniques with genetic and phenotypic identification. Plates with PDA medium containing sampled biological material were incubated at 8 ± 0.5 °C. The density of mycobiota inside the cave ranged from 37.4 to 71 CFU 1 m−3 of air and 63.3 CFU 1 m−3 of air outside the cave. Thus, the level of fungal spores did not exceed the standards for the mycological quality of the air. A total of 18 species were isolated during the study, and some species may be potentially dangerous to people with weakened immune system. All fungal species were present inside the cave and only seven of them were outside. Cladosporium cladosporioides dominated in the external air samples and Mortierella parvispora was cultured most frequently from internal air samples. To our knowledge, this is the first discovery of the fungal species such as Coniothyrium pyrinum, Cystobasidium laryngis, Filobasidium wieringae, Leucosporidium drummii, M. parvispora, Mrakia blollopis, Nakazawaea holstii, and Vishniacozyma victoriae in the air inside the underground sites. Moreover, C. pyrinum, C. laryngis, L. drummii, M. blollopis, and N. holstii have never been detected in any component of the underground ecosystems. There are possible reasons explaining the detection of those species, but global warming is the most likely.
... Fungal rDNA was amplified using the primer ITS1 (5′-TCCGTAGGTGAACCTGCGG-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′). PCR was performed in T100 Thermal Cycler (Bio-Rad), according to Dyląg et al. [22]. The PCR products were verified by electrophoretic separation on a 1.2% agarose gel and, subsequently, purified using Clean-UP (A&A Biotechnology) and sequenced by Macrogen Europe (Netherlands). ...
Houseplants are being grown to increase the aesthetic value of indoor space but also to elevate air quality. Their infections with phytopathogens, however, not only have an impact on the plant physiology and appearance but also may lead to air contamination and, in consequence, affect human health. Present research is a case study of Echinopsis oxygona, known as the Easter Lily cactus, infected with a fungal pathogen. The phenotypic and molecular studies were conducted to identify the etiological agent of the lesions. Colony appearance and growth on various media, as well as the presence of fungal propagation structures were evaluated. Internal transcribed spacer (ITS) sequences from the isolated cultures were obtained, and the BLAST analysis was performed to estimate genetic similarity. The phenotypic and molecular tests allowed to identify the pathogen as Fusarium oxysporum and, to our knowledge, it is the first report on E. oxygona (as a pot plant) infected with this species in Poland. This fungus is a soil-born species and a well-known toxin producer. Therefore, it does not only reduce aesthetic value of the infected plant but also may lead to air contamination with mycotoxins and fungal structures.
... People who cannot prevent the occurrence of the mould problem, they need to resort to chemical agents, special plasters and anti mould paint or major flat renovation. Because fungi are very difficult to get rid of if they have grown deep into various materials, including plasters on the walls and ceilings of the rooms [25,26]. It is also possible to use physical methods to preventing and combating attacks of fungus as well as other harmful living organisms (e.g. ...
The development of indoor molds is mainly connected to the room conditions, like air humidity, temperature or pH. The sanitary rooms, usually with elevated dampness, are a good environment for fungal growth and should be monitored for fungal concentration due to the risk of an elevated exposure to fungal allergens and mycotoxins. In the present work the diversity and the concentrations of fungal pollutants in the dormitory bathroom of Polish University was evaluated for both the ceiling and the air. The results showed that Cladosporium cladosporoides was the only species found on the dark stains on the ceiling, but in the air the variety of fungi was higher, with a total of 6 species: Aspergillus flavus, A. niger, C. cladosporioides, Fusarium poae, Penicillium chrysogenum and Rhizopus nigricans. The increased concentration of A. niger and C. cladosporoides in the air suggests further control of the air pollution in order to avoid excessive exposure to mycotoxins and allergens. Moreover, the concentration of airborne fungi inside the facility exceeded some standards of fungal air contamination stated as dangerous for human health. Our study allows for unambiguously identifying isolated fungi and develop recommendations, to prevent the present outbreak of fungi as well as in the future.
... Plates were incubated in the dark at 15 and 20 ± 1 • C for 7-42 days. The fungi were identified according to available monographs [46][47][48][49][50][51][52][53][54][55][56][57][58]. Additionally, the phenotypes of some fungi were also compared with strains from R. Ogórek's collection (Department of Mycology and Genetics, Institute of Genetics and Microbiology, University of Wrocław, Wrocław, Poland), which were identified using phenotypic and molecular studies, based on their internal transcribed spacer (ITS) region sequences deposited in the National Center for Biotechnology Information (NCBI, Bethesda, Rockville, MD, USA). ...
Bats play important functions in ecosystems and many of them are threatened with extinction. Thus, the monitoring of the health status and prevention of diseases seem to be important aspects of welfare and conservation of these mammals. The main goal of the study was the identification of culturable fungal species colonizing the wing membranes of female greater mouse-eared bat (Myotis myotis) during spring emergence from the “Nietoperek” underground hibernation site by the use of genetic and phenotypic analyses. The study site is situated in Western Poland (52°25′ N, 15°32′ E) and is ranked within the top 10 largest hibernation sites in the European Union. The number of hibernating bats in the winter exceeds 39,000 individuals of 12 species, with M. myotis being the most common one. The wing membranes of M. myotis were sampled using sterile swabs wetted in physiological saline (0.85% NaCl). Potato dextrose agar (PDA) plates were incubated in the dark at 8, 24 and 36 ± 1 °C for 3 up to 42 days. All fungi isolated from the surface of wing membranes were assigned to 17 distinct fungal isolates belonging to 17 fungal species. Penicillium chrysogenum was the most frequently isolated species. Some of these fungal species might have a pathogenic potential for bats and other mammals. However, taking into account habitat preferences and the life cycle of bats, it can be assumed that some fungi were accidentally obtained from the surface of vegetation during early spring activity. Moreover, Pseudogymnoascus destructans (Pd)—the causative agent of the White Nose Syndrome (WNS)—was not found during testing, despite it was found very often in M. myotis during previous studies in this same location.
... lobatumVanderwolf et al. (2013) Chr. merdariumVanderwolf et al. (2013),Dyląg et al. (2019) Chr. pallidum Y YChr. ...
... al. (2013, 2014a, b),Vanderwolf et al. (2013),Kokurewicz et al. (2016),Pusz et al. (2018a),Dyląg et al. (2019) Sa. zeaeMitova et al. Ogórek et al. (2013, 2014a,Pusz et al. (2018a) Sclerotinia sp.Vanderwolf et al. (2013) Scolecobasidium Scolecobasidium anelliiVanderwolf et al. (2013) Sc. anomalumVanderwolf et al. (2013) Sc. constrictumVanderwolf et al. (2013) Sc. lascauxenseVanderwolf et al. (2013), Martin-Sanchez et al. (2014), Pfendler et al. (2019) Sc. terreum Vanderwolf et al. (2013) Scopulariopsis Scopulariopsis asperula Y Y Vanderwolf et al. (2013) Sco. ...
Karst caves are characterized by darkness, low temperature, high humidity, and oligotrophic organisms due to its relatively closed and strongly zonal environments. Up to now, 1626 species in 644 genera of fungi have been reported from caves and mines worldwide. In this study, we investigated the culturable mycobiota in karst caves in southwest China. In total, 251 samples from thirteen caves were collected and 2344 fungal strains were isolated using dilution plate method. Preliminary ITS analyses showed that these strains belonged to 610 species in 253 genera. Among these species, 88.0% belonged to Ascomycota, 8.0% Basidiomycota, 1.9% Mortierellomycota, 1.9% Mucoromycota, and 0.2% Glomeromycota. The majority of these species have been previously known from other environments, and some of them are known as mycorrhizal or pathogenic fungi. About 52.8% of these species were discovered for the first time in karst caves. Based on morphological and phylogenetic distinctions, 33 new species were identified and described in this paper. Meanwhile, one new genus of Cordycipitaceae, Gamszarea, and five new combinations are established. This work further demonstrated that Karst caves encompass a high fungal diversity, including a number of previously unknown species.
Taxonomic novelties: New genus: Gamszarea Z.F. Zhang & L. Cai; Novel species: Amphichorda cavernicola, Aspergillus limoniformis, Aspergillus phialiformis, Aspergillus phialosimplex, Auxarthron chinense, Auxarthron guangxiense, Auxarthronopsis globiasca, Auxarthronopsis pedicellaris, Auxarthronopsis pulverea, Auxarthronopsis stercicola, Chrysosporium pallidum, Gamszarea humicola, Gamszarea lunata, Gamszarea microspora, Gymnoascus flavus, Jattaea reniformis, Lecanicillium magnisporum, Microascus collaris, Microascus levis, Microascus sparsimycelialis, Microascus superficialis, Microascus trigonus, Nigrospora globosa, Paracremonium apiculatum, Paracremonium ellipsoideum, Paraphaeosphaeria hydei, Pseudoscopulariopsis asperispora, Setophaeosphaeria microspora, Simplicillium album, Simplicillium humicola, Wardomycopsis dolichi, Wardomycopsis ellipsoconidiophora, Wardomycopsis fusca; New combinations: Gamszarea indonesiaca (Kurihara & Sukarno) Z.F. Zhang & L. Cai, Gamszarea kalimantanensis (Kurihara & Sukarno) Z.F. Zhang & L. Cai, Gamszarea restricta (Hubka, Kubátová, Nonaka, Čmoková & Řehulka) Z.F. Zhang & L. Cai, Gamszarea testudinea (Hubka, Kubátová, Nonaka, Čmoková & Řehulka) Z.F. Zhang & L. Cai, Gamszarea wallacei (H.C. Evans) Z.F. Zhang & L. Cai.
... Plates with fungi were cultured at 23 ± 1.0 • C for 5-14 days. The observations were analyzed according to available monographs [33][34][35][36][37][38][39][40][41][42][43]. Microscopic slides were dyed with lactophenol cotton blue (LPCB), Sigma, Saint Louis, MO, USA), and photographs were taken with an Axio Image.M1 (Zeiss, Göttingen, Germany). ...
The Orchidaceae family is a diverse family of flowering plants that occur naturally in most parts of the world. However, fungal communities inhabiting different parts of orchids are not sufficiently described. The aim of the study was to conduct a mycological evaluation of Epipactis helleborine and E. purpurata (Orchidaceae), which grow naturally in Lower Silesia (SW Poland), by identifying the species composition of the culturable micromycetes fungi on the surfaces of the plants and from the inner layers of the tissues. Fungi were identified based on a phenotypic and genotypic analysis. To our knowledge, this is the first such analysis. This study showed that more species of micromycetes were cultured from E. helleborine compared with E. purpurata. The flowering plants of E. helleborine were inhabited by the largest number of culturable fungal species (13 species), and the fewest species were isolated from the flowering plants of E. purpurata (eight species). Some of these fungal species may be pathogens of the plants. The surface tissues of the orchids were mainly inhabited by Mucor moelleri and/or Penicillium biourgeianum. The inner layers of these plants were the most colonized by Alternaria tenuissima and/or Arthrinium arundinis and/or Fusarium sporotrichioides. The relative dominance of these fungal species depended mainly on the development phase of the plants.
Soil and sediment host microorganisms are able to survive in extremely resource-limited environments. Therefore, more and more attention is being paid to cave sediments as a reservoir of microbiota. The aim of this study is the speleomycological evaluation of the culturable soil and sediment fungal communities in the Brestovská Cave. To explore the origins of fungi, speleomycological studies were conducted both inside and outside the cave under investigation. Additionally, two incubation temperatures (5 and 24 °C) were used to increase the species spectrum of isolated fungi. To achieve the most accurate species identification, we combined an assessment of morphological characteristics of the isolates with molecular sequencing (ITS, internal transcribed spacer). Twenty different species were found and the most frequent was Penicillium commune, fol-lowed by Trichosporiella cerebriformis and Pseudogymnoascus pannorum. To our knowledge, our study has enabled the first identification of fungal species such as Penicillium swiecicki, Cephalotrichum hinnuleum, Cosmpospora berkeleyana, Lecythophora hoffmannii, Ambomucor seriatoinflatus, and Mortierella minutissima in underground sites. Our data showed that the abundance and composition of the fungal community varied between the indoor and outdoor samples and thus from the entrance and less visited sites deeper in the cave.
There is increasing concern about exposure to airborne fungi. Research on mycotoxins shows that just as airborne fungal spores are everywhere in our environments, so too are their metabolic products. Mycotoxins with their cytotoxic, genotoxic, mutagenic, and teratogenic properties are persistent threats to human and animal health. Assessment of fungal exposure is notoriously challenging due to the numerous factors that contribute to the variation of fungal concentrations in environments. This paper reviews the homogeneity in the fungal species composition of these bioaerosols on a large geographical scale and the different drivers that shape these fungal communities which still remain unclear yet seem to be strongly governed by geographical location.
Abstract
Poly (thiourea–formaldehyde) resin (PTUFR) is synthesized by gamma rays. Fourier transform infrared (FTIR), 1H-NMR and 13C-NMR spectroscopy, transmission electron microscopy (TEM), and gel permeation chromatography (GPC) were the techniques used to characterize PTUFR. The results confirmed the formation of PTUFR. The antimicrobial activity of PTUFR was tested against the most resistant isolated aquatic microbes Gram-positive bacteria (Bacillus subtilis), Gram-negative bacteria (Pseudomonas aeruginosa), and fungi(Candida albicans). Through the interaction with internal cellular components and cell membranes, PTUFR showed potential for bio-application in inactivating microbial cells by killing them. The two cell death mechanisms of the reactions of PTUFR with the proteins of the cell wall led to reduced permeability, as well as reactive oxygen species (ROS) with high release rates from PTUFR, which in a row caused damage to DNA and cellular components. Furthermore, the cytotoxicity of PTUFR against human breast cancer MCF-7 and HepG2 liver cancer cells was studied. The cytotoxicity of PTUFR resulted in the inhibition of cell growth on the lines of the cancer cells. [Z1] The exposure of MCF-7 and HepG2 cancer cells to PTUFR for 48h displayed a potential apoptotic activity by significant upregulation of p53 gene expression and cytotoxic activity by downregulation of Bcl-2 gene expression. These results may also contribute to an unconventional strategy for environmental and medical applications, such as safe wastewater disinfection and cytotoxic activity.
