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Fungi in Public Heritage Buildings in Poland

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Fungi in Public Heritage Buildings in Poland Pol. J. Environ. Stud. Vol. 29, No. 5 (2020), 3651-3662 DOI: 10.15244/pjoes/112213 ONLINE PUBLICATION DATE: 2020-04-07 Original Research Fungi in Public Heritage Buildings in Poland Hanna Kwaśna1*, Alicja Kuberka2 1Department of Forest Pathology, Faculty of Forestry, Poznań University of Life Sciences, Poznań, Poland 2Faculty of Wood Technology, Poznań University of Life Sciences, Poznań, Poland Received: 10 June 2019 Accepted: 8 September 2019 Abstract With the prevalence of asthma increasing worldwide in recent decades there is a growing need to determine the diversity and spatiotemporal dynamics of airborne microbes in indoor environments. Hence, the aims of this study were to detect and identify fungi present in public heritage buildings (libraries, museums, churches) in Poland. Samples of indoor dust and biofilm were collected from 60 areas in 10 such buildings (mostly in urban environments) in different seasons in 2008-2011. Mycological analysis was based on the culture-dependent method using 2% MEA and SNA. In 300 samples there were at least 160 fungal species belonging to more than 70 genera. The most common were Acremonium, Alternaria, Arthrinium, Aspergillus, Aureobasidium, Botrytis, Chaetomium, Cladosporium, Fusarium, Monodictys, Mucor, Nectria, Oidiodendron, Penicillium, Phoma, Rhizopus, Sarocladium, Simplicillium, Scopulariopsis, Talaromyces, Trichoderma and Tritirachium. Apart from filamentous fungi and yeasts Candida( and Sporobolomyces), single species of Oomycota (Pythium insidiosum), Actinobacteria and Myxogastia were detected. Most of the taxa recorded have been regarded as important indoor contaminants. Potential health hazards and the deterioration of contaminated objects are discussed. Keywords: fungi, deterioration, church, museum, national heritage Introduction activity and their ability to grow at low water activity (aw) values allow them to inhabit wood, paper, Indoor microbial communities are a ubiquitous paints, glues, soil, dust and surfaces consisting of part of buildings. They accumulate in the air and inorganic moist material (e.g. glass, fiberglass, metal dust. They are affected by the microbial content of the or concrete) covered with dust, air contaminants air outdoors. Dust in indoor air is a complex mixture or even fingerprints, which create an invisible layer of particles from soil, plants (fragments, pollen) and of biofilm. During growth they produce and excrete animals, but also from construction materials, furniture many strong enzymes and acids that can efficiently and exhibited or stored objects. destroy and/or disintegrate organic and inorganic The majority of fungi present in the indoor materials. environment is saprotrophic. Their enormous enzymatic Microbial particles adhere to solid surfaces by surface hydrophobicity and electrostatic forces or by specific interactions between cell-surface molecules (i.e. hydrophobins and glycoproteins), and the surface *e-mail: [email protected] of an object. Strong, irreversible conidial adhesion 3652 Kwaśna H., Kuberka A. generally depends on spore hydration and specific There are four levels of biocontainment precaution activation, or the expression of surface adhesion required for contact with biological agents, including molecules such as extracellular matrix components that microorganisms (biosafety level 1-4). Risk groups act as glue attaching the cell to the surface. BSL–1, BSL–2, BSL–3 and BSL–4 contain biological Biofilms can form on any surface after being agents that pose, respectively, low, moderate, serious conditioned by proteins and other molecules. After and very serious risk to humans and the environment. attachment, microbes begin to release polysaccharides, With the prevalence of asthma increasing worldwide proteins and DNA, which help them to stick better in recent decades there is a growing need to determine and provide protection from UV light or disinfectants. the diversity and spatiotemporal dynamics of airborne Eventually, the biofilm matures, and can then be microbes in the indoor environment. Hence, the aim of disrupted or dried out, its fragments becoming this study was to detect, identify and quantify the fungi aerosolized in the air and deposited in dust. present in public heritage buildings (libraries, museums, Fungal fragments in dust vary in size from <1 μm to churches) in Poland. Their potential health hazard is >100 μm. Size and shape contribute to their differential discussed. dispersal, deposition and resuspension in the indoor environment as well as to the induction of respiratory health issues in humans. Smaller particles (e.g. bacteria, Materials and Methods ascomycetous microconidia and fungal fragments) mix more efficiently in the entire room space and stay Samples were collected in 10 libraries, museums and airborne longer, or even constantly [1]. churches at 10 locations in Poland, in different seasons, The total concentration of fungi in dust can vary in 2008-2011. greatly. Mean concentrations of between 104 and Dust from the air was actively sampled using a 105 CFU g−1 are typically reported from culture- dust stream collector, first sterilized by soaking in 10% dependent studies. Total fungal cell counts obtained bleach and drying under UV radiation, attached to a using the qPCR assays have been reported to be at least domestic vacuum cleaner. Each sample was filtered 10- to 100-fold higher. Total cell equivalent (CE) counts through a 2-mm sieve for 10 min and refrigerated at can be 3.9×105 – 1.4×109 CE g−1 and corresponding 4ºC (if not processed immediately). For active sampling viable counts 6.1×104 – 2.1×107 CFU g−1 [2]. Levels may from objects, micro-fragments of biofilm were collected vary from non-detectable to very high depending on by sterile scalpel, sterile swabs or H+ Nylon Amersham- geographical location, season, type of building, type of membranes. These were placed in contact with the sample and external factors, as well as human activities surfaces being sampled and pressed for 30 s. In the such as cleaning. Resuspension of particles from laboratory, the sieves, swabs or nylon membranes were surfaces in indoor environments is mainly influenced cut into 5 x 5 mm pieces. Thirty-six pieces from each by human activity. Fungal hyphal fragments are the sample were put onto 2% malt extract agar (MEA; 20 g first to lose their viability, i.e. the ability to start new Difco powdered malt extract, 1 g peptone, 20 g glucose, growth. Dispersal spores persist longer and may stay 20 g agar, 1 l distilled water) and synthetic nutrient agar viable in dust over decades. (SNA; 0.2 g glucose, 0.2 g sucrose, 1 g KH2PO4, 1 g Fungal spores and fragments carry many KNO3, 0.5 g MgSO4.7H2O, 0.5 g KCl, 15 g agar, 0.1 g compounds with the potential to affect human health. streptomycin sulphate, 1 l distilled water) in six Petri These include structural compounds such as ergosterol dishes (9 cm diam., six pieces in each Petri dish) and and (1 → 3) – beta-D-glucan – common to all fungi, incubated at 25ºC for 5-25 days. enzymes and other proteins with less universal The morphology of fungal colonies was observed by presence, as well as nonvolatile metabolic products such stereomicroscope (Zeiss, 10-20x) and fungal structures, as mycotoxins, and volatile compounds (VOCs) such mainly conidiophores and conidia, stained with Lugol’s as alcohols, terpenes, and aldehydes, the production of iodine, by optical microscope (Zeiss, 40-100x ). Fungi which is usually restricted to a few individual fungal were identified according to mycological keys, with tax- species or strains [3]. on omy up dated ac cord ing to the CBS Fun gal Bio di ver- The first studies on fungal density and diversity in sity Cen tre nomen cla ture. The frequency of a fungus in indoor dust were made in the 1940s and 1950s. Health a sample was defined as the percentage of sample pieces effects of exposure to fungi have been presented in the (inocula) colonized. literature [4]. The exposure to fungal species such as Alternaria, Aspergillus, Aureobasidium and Penicillium in humans may induce illness such as rhinitis, Results sinusitis, asthma or alveolitis. Fungal bioaerosol, consisting of spores, hyphal fragments and volatile In 300 samples of indoor dust and biofilm collected metabolites may cause serious respiratory infections in in 60 areas of 10 libraries, museums and churches immunocompromised individuals, bronchial irritation (mostly in urban environments) in Poland there were and allergy. Both single substances and synergistic at least 160 fungal species belonging to more than effects contribute to toxicity. 70 genera (Table 1). Fungi inPublicHeritageBuildings... Table 1. Fungi in dust and biofilm on objects in libraries, museums and churches in Poland No. Taxon Location1 Character Pathogenicity Oomycota A B C D E F G H I J 1. Pythium insidiosum De Cock, L. Mend., A.A. Padhye, Ajello & Kaufman animal and human pathogens BSL 22 13 Fungi 1. Absidia cylindrospora Hagem saprotrophs, possible human patho- GRAS 2 2. Absidia repens Tiegh. gens GRAS 1-2 3. Acremonium blochii (Matr.) W. Gams animal and human pathogens NC 1-4 1 4. Acremonium fusidioides (Nicot) W. Gams plant pathogen GRAS 1 5. Acremonium hyalinulum (Sacc.) W. Gams animal pathogen NC 1 6. Alternaria alternata (Fr.) Keissl. BSL 1 1-4 2-4 1 2 4 1 4 3 7. Alternaria atra (Preuss) Woudenb. & Crous BSL 1 4 8. Alternaria chartarum Preuss saprotrophs, plant and human BSL 1 1 2 1 4 1 9. Alternaria infectoria E.G. Simmons pathogens BSL 1 1-2 10. Alternaria oudemansii (E.G. Simmons) Woudenb. & Crous BSL 1 2 4 1 2 11. Alternaria tenuissima (Kunze) Wiltshire BSL 1 2 12. Arthrinium arundinis (Corda) Dyko & B. Sutton NC 2 2 saprotrophs, human pathogens 13. Arthrinium phaeospermum (Corda) M.B. Ellis BSL 1 1 1 1 1 14. Ascomycota sp. 2 15. Ascotricha chartarum Berk. potential human pathogen NC 1 16. Aspergillus caesiellus Saito NC 1 17. Aspergillus clavatonanicus Bat., H. Maia & Alecrim NC 1 18. Aspergillus deflectus Fennell & Raper NC 1 1 19. Aspergillus glaucus (L.) Link BSL 1 1 1 1 20.
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