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Abstract The different organisms, ranging from plants to bacteria, and viruses that dwell on built cultural heritage can be passive or active participants in conservation processes. For the active participants, particular attention is generally given to organisms that play a positive role in bioprotection, bioprecipitation, bioconsolidation, bioremediation, biocleaning, and biological control and to those involved in providing ecosystem services, such as reducing temperature, pollution, and noise in urban areas. The organisms can also evolve or mutate in response to changes, becoming tolerant and resistant to biocidal treatments or acquiring certain capacities, such as water repellency or resistance to ultraviolet radiation. Our understanding of the capacities and roles of these active organisms is constantly evolving as bioprotection/biodeterioration, and biotreatment studies are conducted and new techniques for characterizing species are developed. This brief review article aims to shed light on interesting research that has been abandoned as well as on recent (some ongoing) studies opening up new scopes of research involving a wide variety of organisms and viruses, which are likely to receive more attention in the coming years. Key points • Organisms and viruses can be active or passive players in heritage conservation • Biotreatment and ecosystem service studies involving organisms and viruses are shown • Green deal, health, ecosystem services, and global change may shape future research

Biocleaning of cultural heritage items is mainly performed using living microorganisms. Approaches utilizing the enzymes of isolated microorganisms have not been frequently investigated. To find an enzymatic alternative for the removal of an oil-based overpainting, we focused on the characterization and use of a yeast Extracellular Enzymatic Mixture (EEM). A historical silk yeast was selected for its lipolytic properties and its EEM was extracted after cultivation on a medium supplemented with linseed oil. The EEM protein content was visualized by SDS-PAGE, its concentration assessed by fluorimeter and the enzymatic activity evaluated by p-NPP spectrophotometric lipase assay. The yeast growth was suppressed by adding diverse metal ions (Cd, Zn, Cr and Cu) in Reasoner's 2A (R2A) broth, while the quantity and activity of EEM were affected by adding Fe and Pb. Various delivery systems (agar-agar, tylose and klucel G) alone or in a combination with EEM were assayed on the historical painting surface. The colorimetric measurements and the ATR-FTIR analysis indicated that the combinations tylose-EEM and klucel G-EEM can be easily and effectively applied as biocleaning procedures to remove oil-based overpainting from fragile and valuable historical painting surfaces.

The catabolism of milk protein in cheese is one way how the microorganisms influence the sensorial characteristics of the final product. In this investigation, we paid attention to four genes [prtP (cell-envelope proteinase gene), pepX (X-prolyl dipeptidyl aminopeptidase gene), pepN (aminopeptidase gene) and bcaT (branched chain aminotransferase gene)] responsible for the production of volatile aroma-active compounds from milk proteins by lactic acid bacteria (LAB). We studied the dynamics of these genes and their corresponding LAB host, during the maturation of a raw ewes' milk-based cheese, using metagenomics and metatranscriptomics approaches. The transcriptome-oriented experiments included the analysis of total RNA (at three stages of cheese maturation) and also the construction of specific cDNA sub-libraries of the abovementioned genes. The proteolytic transcriptome analysis was supported by following the transcription activity of 16S rRNA gene and by metagenomic investigation. The combination of the described methods permitted to screen the dynamics of targeted genes throughout the cheese production. Lactococci were the major players in the LAB group, but the analysis provided also information on the role and properties of members of the genus Lactobacillus, such as Lb. rhamnosus, Lb. helveticus, Lb. pentosus, Lb. curvatus, Lb. parabuchneri, Lb. plantarum, Lb. brevis, Lb. delbrueckii, Lb. paracasei, Lb. fermentum and Lb. heilongjiangensis, proteolysis-related genes of which were active during cheese ripening.

Distribution and biodiversity of soil microscopic fungi in 5 areas of old environmental loads in Slovakia were studied in relation to very low amount of organic matter (%TOC from 0.2 to 3.54) and to the pH gradient from ultra-acidic ( 3.5) to very strongly alkaline ( 9.0). All soil samples were affected by several hundred years of mining activities and contained heavy metals and other toxic elements: arsenic, cadmium, copper, zinc, antimony, lead. Concentrations of toxicants highly exceeded their limited values. Fifty-three genera and 112 species of microscopic fungi were identified. Among them, Zygomycota occurred very rarely (8 genera and 12 species), except of samples with the highest content of TOC (2.01-3.54% - samples 2 and 6), regardless their pH. Though, on the other hand, from some similar samples (3, 5 and 9), incl. those with relatively high TOC (0.14-2.62%), the lower fungi were not recovered. Forty one genera and 95 species of Ascomycota represented the most abundant fungal phylum in all investigated samples. Among them, Penicillium chrysogenum var. chrysogenum, Aspergillus niger and Neosartorya fischeri were isolated the most often. Phytopathogenic moulds of Bionectria ochroleuca, Lewia infectoria, Phoma macrostoma and Phlebia acerina were also occurred frequently. The highest biodiversity of microfungal community was recorded in the extreme acidic environment, followed by the neutral, ultra-acidic and the very strong acidic ones. There was no similarity in microfungal spectrum found in the samples studied. Except of the ultra acidic and extreme acidic samples (1-2) as well as the ultra acidic and strong acidic ones (1-4) with the most rich mycobiota, that may indicate a certain similarity degree.

Living microorganisms have been used for biocleaning of cultural heritage items, but the application of only extracellular enzyme mixtures (EEMs) from isolated bacteria has not previously been considered for biorestoration work. In this investigation, the proteolytic characteristics of the EEMS from three bacterial isolates (Paracoccus sp., Bacillus flexus and Exiguobacterium undae) were determined and their extent of inhibition in the presence of the proteolytic inhibitors 1,10-phenanthroline monohydrate, EDTA, leupeptin, bestatin and various metal ions was examined. These EEMs were used to remove the animal glue present on the spine of bound newspapers. FT-IR analysis showed that animal glue was effectively removed by a solution of tylose and EEM. The EEM of E. undae gave the best results. Our study describes a simple strategy for the production and characterization of enzymatic mixtures targeted to the degradation of a specific substrate and their subsequent use for biocleaning historical objects.

The potential of K2FeO4 as a pretreatment agent of a lignocellulosic material was examined on leaves of Acer platanodides as the sole substrate for biogas production by anaerobic digestion carried out through modelling laboratory-scaled semi-continuous reactors differing in loading rates and substrate (pretreated and untreated leaves). The quality of bioagas produced by K2FeO4-pretreated leaves was significantly better in terms of higher methane content and lower content of H2S. K2FeO4 had no crucial influence on growth inhibition of biogas-producing bacteria, which were analysed by comprehensive culture-independent methods utilising high-throughput sequencing of specific genes [bacterial and archaeal 16S rRNA, formyltetrahydrofolate synthetase gene (fhs), methyl-coenzyme M reductase α subunit gene (mcrA) and fungal internal transcribed spacers (ITS)]. The higher amount of CH4 in biogas utilising pretreated leaves as substrate could be caused by a shift to acetoclastic methanogenesis pathway, which was indicated by the higher amount of homoacetogenic bacteria and acetotrophic methanogens detected in those reactors.

A historical crypt offers us a particular view of the conditions of some buried materials (in this case textiles) and the various biogenic phenomena to which they were subjected over the centuries. In addition, significant knowledge can come by studying the DNA of buried objects which allows the recognition of materials, but also to reveal some practice of the funeral ceremony. In this study, the deteriorating microbial communities colonizing various funeral textile items were identified and characterized using microscopic observation, cultivation, polymerase chain reaction (PCR) and sequencing, hydrolytic tests, and culture-independent analysis (high-throughput sequencing, MinION platform). Different PCR assays and consequent sequencing of amplicons were employed to recognize the animal origin of bodice reinforcements and the type of plant used to embellish the young girl. The analysis of ancient DNA (aDNA from animal and plant) was also completed by the application of high-throughput sequencing through Illumina platform. The combination of all these techniques permitted the identification of a complex microbiota composed by dangerous degradative microorganisms able to hydrolyze various organic substrates such as fibroin, keratin, and cellulose. Bacteria responsible for metal corrosion and bio-mineralization, and entomopathogenic and phytopathogenic fungi. The analysis of aDNA identified the animal component used in bodice manufacturing, the plant utilized as ornament and probably the season of this fatal event.

This microbiological survey was performed to determine the conservation state of a mummy in the Slovak castle of Krasna Horka and its surrounding environment. Culture-dependent identification was coupled with biodegradation assays on keratin, gelatin and cellulose. Next Generation Sequencing (NGS) using Illumina platform was used for a deeper microbial investigation. Three environmental samples were collected: from the glass of the sarcophagus, from the air inside it, and from the air of the chapel where the mummy is located. Seven different samples were taken from mummy's surface: from the left ear, left-hand palm, left-hand nail, left instep, right hand, abdomen and mineral crystals embedded within the skin. Three internal organ samples, from the lung, pleura and stomach, were also included in this study. Together, the culture-dependent and culture-independent analyses revealed that the bacterial communities present had fewer taxa than the fungal ones. The mycobiome showed the largest variability and included Epicoccum nigrum, Penicillium spp., Alternaria spp., Aspergillus spp., Cladosporium spp. and Aureobasidium pullulans; many other Ascomycota and Basidiomycota genera were detected by NGS. The most interesting results came from the skin mineral crystals and the internal organs. The hydrolytic assays revealed those microorganisms which might be considered dangerous 'mummy pathogens'. © 2018 Society for Applied Microbiology and John Wiley & Sons Ltd.

Different identification strategies employing new methodologies, namely, culture-dependent (using matrix-assisted laser desorption/ionization – time-of-flight mass spectrometry, MALDI-TOF MS) and culture-independent (using high-throughput sequencing on Illumina MiSeq platform) approaches, were applied for the first time to analysis of the deteriorating microflora of book samples. We compared two different identification techniques coupled to microbial cultivation: DNA sequencing and MALDI-TOF MS. The two identification systems produced almost the same results. DNA sequencing was able to better identify all the recovered isolates, while MALDI-TOF MS failed to properly recognize Streptomyces ambofaciens and Lysinibacillus fusiformis, the latter method permitting identification at least at genus level. In addition, the isolates Myxotrichum deflexum and Oidiodendron cerealis were not identified by the MALDI-TOF MS approach. This incongruence was caused by the absence of different kinds of microorganisms in the MALDI BioTyper reference database. The Illumina high-throughput sequencing coupled to non-invasive sampling allowed the identification of a complex bacterial and fungal community. The study presents the satisfactory performance of two novel alternative identification methods (MALDI-TOF MS and Illumina MiSeq) for the investigation of microflora colonizing archival items.

In this work, we describe the preparation and characterization of a biopreparate for efficient and rapid animal glue removal. The biopreparate is based on the extracellular proteolytic enzymes of an Exiguobacterium undae environmental isolate. Liquid chromatography-mass spectrometry analysis showed that the biopreparate is predominantly composed of hydrolytic enzymes-proteases and peptidases, nucleases, peptide ABC transporter substrate-binding proteins, and a phosphatase. The two main proteins present are bacillolysin and a peptide ABC transporter substrate-binding protein. Inhibition and proteomic analyses of the biopreparate revealed that bacillolysin, a neutral metalloendopeptidase, is mainly responsible for its proteolytic activity. This biopreparate was able to satisfactorily remove two types of animal glue from different kinds of material surfaces. These results suggest that this biopreparate could serve as a potential new tool for the restoration of historical objects rather than living microorganisms.

Culture-dependent and culture-independent strategies were applied to investigate the microbiota of autumn undamaged and damaged berries, winter berries and ice wine must samples of Grüner Veltliner (Veltlínske zelené) from Small Carpathian wine-producing region. One hundred twenty-six yeasts and 242 bacterial strains isolated from several microbiological media (YPD, PDA, R2A, GYC, MRS and MRS-T) were clustered by ITS-PCR and subsequent Qiaxcel electrophoresis. Representatives of each cluster were identified by sequencing. The extracellular hydrolytic properties and intracellular activities of esterase and β-glucosidase of isolates were assayed. The culture-independent approach permitted the analysis of extracted DNA and RNA coupling DGGE fingerprinting with construction of clone libraries (bacterial and fungal; DGGE-cloning). The combination of the two approaches provided comprehensive data that evidenced the presence of a complex microbiota in each analyzed sample. RNA and DNA analyses facilitated differentiation of living microorganisms from the entire microbiota. Diverse microbial communities colonized the autumn and winter berries. Generally, the combination of results obtained by the methods suggested that the must samples contained mainly Saccharomyces cerevisiae, Metschnikowia spp., Hanseniaspora uvarum, Lactococcus lactis and Leuconostoc spp. The strains exhibited interesting esterase and β-glucosidase properties, which are important for aroma formation in wine. Fermentation strategies utilising these microorganisms, could be attempted in the future in order to modulate the ice wine characteristics.

Six essential oils (from oregano, thyme, clove, lavender, clary sage, and arborvitae) exhibited different antibacterial and antifungal properties. Antimicrobial activity was shown against pathogenic (Escherichia coli, Salmonella typhimurium, Yersinia enterocolitica, Staphylococcus aureus, Listeria monocytogenes, and Enterococcus faecalis) and environmental bacteria (Bacillus cereus, Arthrobacter protophormiae, Pseudomonas fragi) and fungi (Chaetomium globosum, Penicillium chrysogenum, Cladosporium cladosporoides, Alternaria alternata, and Aspergillus fumigatus). Oregano, thyme, clove and arborvitae showed very strong antibacterial activity against all tested strains at both full strength and reduced concentrations. These essential oils showed different fungistatic and fungicidal activities when tested by direct application and in the vapor phase. The genotoxic effects of these oils on HEL 12469 human embryo lung cells were evaluated using an alkaline comet assay for the first time, revealing that none of the oils induced significant DNA damage in vitro after 24 h. This study provides novel approaches for assessing the antimicrobial potential of essential oils in both direct contact and the vapor phase and also demonstrates the valuable properties of the phenol-free arborvitae oil. These results suggest that all the tested essential oils might be used as broad-spectrum anti-microbial agents for decontaminating an indoor environment.

Wooden pipe organs are precious objects of cultural heritage the degradation of which has been very poorly studied. Fungal strains recovered from an organ dating back to the 19th century included Alternaria mali , Eurotium cristatum , Aspergillus amstelodami , Penicillium crustosum and Aspergillus sydowii . These isolates were used to artificially contaminate wooden pipes treated with linseed oil varnish and varnish-free. The pipes were incubated at different temperatures and relative humidity for one year. The pipes, incubated at a high relative humidity (70–75%), collapsed due to the biodegradation of glued joints. After one year, inoculated strains ( Alternaria mali , Penicillium crustosum and Aspergillus sydowii ) as well as some non-inoculated Talaromyces rugulosus, Paecilomyces formosus , Cladosporium cladosporioides , Aspergillus versicolor and Epicoccum nigrum were recovered from the pipes. Few strains exhibited lignolytic activity, while almost all were able to hydrolyze the rabbit glue used to join the wood elements of the pipes. A newly developed portable pipe-sound controller did not detect differences of sound spectra between the uncontaminated and the few surviving contaminated pipes. In this study we have demonstrated that glue degradation was critical for pipe preservation and it can be tremendously accelerated above a certain relative humidity threshold. Therefore, it is important to apply a monitoring strategy able to check the environmental conditions, microbial contamination and pipe sound alteration in order to preserve these valuable and rare cultural heritage objects.

In this study the biodegradation characteristics of aromatic aliphatic copolyester Ecoflex was assayed with five different bacterial strains isolated from different kinds of environments and living at diverse optimum of temperature. The biodegradation properties of selected strains (Brevibacterium luteolum, Bacillus thuringiensis, Aeromonas media, Geobacillus kaustophilus and Serratia sp.) were assessed through several plate assays oriented to their proteolytic, lipolytic, esterolytic and cellulolytic abilities; catalase activities were measured too. In addition the bacterial hydrolytic abilities for polyhydroxybutyrate, polylactic acid and polybutyrate adipate terephthalate were obtained using specific culture media including also a specific Ecoflex agar. The biodegradation dynamics of selected strains were evaluated mainly by the incubation of Ecoflex foils with a specific bacterium during a period from 3 days until 16 weeks and the change of the degradability of the Ecoflex surface in nitrogen has been assessed by nonisothermal chemiluminescence measurements from 40 to 250 °C. Scanning electron microscope observation of inoculated foils was applied too. The comparison of the results showed the complexity of biodegradation phenomena. The concentration of oxygenated products measured by reached the maximum after 2 weeks of cultivation for strains G. kaustophilus and B. thurigiensis, after 4 weeks of cultivation for strain B. luteolum, and 8 weeks of cultivation for strains Serratia sp. and A. media. Each strain evidenced its own hydrolytic optimum at different periods, but almost all tested strains demonstrated a decline of hydrolysis up to 16 weeks of incubation. This study demonstrated that nonisothermal chemiluminescence examining momentaneous oxidation state of the polymer surface is a valuable technique to follow the biodegradation dynamics on polymeric film, while the Ecoflex agar is able to select useful polymer-degrading bacteria.

Technosols or technogenic substrates contaminated by potentially toxic elements as a result of iron mining causes not only contamination of the surrounding ecosystem but may also lead to changes of the extent, abundance, structure and activity of soil microbial community. Microbial biomass were significantly inhibited mainly by exceeding limits of potentially toxic metals as arsenic (in the range of 343–511 mg/kg), copper (in the range of 7980–9227 mg/kg), manganese (in the range of 2417–2670 mg/kg), alkaline and strong alkaline pH conditions and minimal contents of organic nutrients. All of the 14 bacterial isolates, belonged to 4 bacterial phyla, Actinobacteria, Firmicutes; β- and γ-Proteobacteria. Thirteen genera and 20 species of microscopic filamentous fungi were recovered. The most frequently found species belonged to genera Aspergillus (A. clavatus, A. niger, A. flavus, A. versicolor, Aspergillus sp.) with the dominating A. niger in all samples, and Penicillium (P. canescens, P. chrysogenum, P. spinulosum, Penicillium sp.). Fungal plant pathogens occurred in all surface samples. These included Bjerkandera adustata, Bionectria ochloleuca with anamorph state Clonostachys pseudochloleuca, Lewia infectoria, Phoma macrostoma and Rhizoctonia sp., Highlights • Alkaline and strongly alkaline Technosol contaminated by As, Cu, Mn, Pb, Zn caused changes in autochtonous microbiota. • Variovorax pantotropha and Staphylococcus pasteuri Sp-12 were isolated. • 20 fungal species belonged to 13 genera were recovered. • The isolated microorganisms showed their high adaptation abilities. • Different isolates can be useful for various industrial and remediation processes.

The research aim was to determine the effectiveness of disinfection methods: thyme essential oil microatmosphere (TEO), silver nanoparticles misting (AgNPs), and low temperature plasma (LTP) by culture-dependent method and RNA analysis. In addition, we examined the influence of disinfection on mechanical and optical properties of paper from historical books with different levels of microbial contamination. All disinfection methods were generally bacteriostatic and fungistatic. The AgNPs misting method was more effective for bacterial inhibition ( R = 60–100%), than LTP ( R = 25–100%) and TEO ( R = 12–100%); all tested methods were less effective for fungi ( R = 0–99.8%). TEO exhibits a broader spectrum of fungicidal activity compared to AgNPs and LTP. The antimicrobial effectiveness depended on the area from where the microorganisms were isolated and their sensitivity to the active agent. A higher disinfection effectiveness was observed for books with higher levels of microbial contamination. RNA concentration was a good marker of antimicrobial activity of disinfection. RNA amount decreased by 95–100% after disinfection with LTP and TEO, and by 29–89% after AgNPs disinfection. Disinfection of paper from historical books with LTP, AgNPs and TEO did not significantly influence or have a positive effect on the mechanical and optical properties of paper from tested historical books. We show that LTP, TEO, and AgNPs can be used as microbiostatics, alternatively to the currently available methods.

A multi-disciplinary investigation was undertaken to determine the cause of corrosion of iron nails from a skeletal structure in a museum setting. Corroded iron nails were removed from a large whale skeleton, that had been assembled in the 1930s and exhibited for over 80 years at the Natural History Museum in London, United Kingdom. Oxford Nanopore Technologies sequencing, SEM-EDS imaging and chemical mapping were used to document the fungal and bacterial communities associated with the corrosion of the iron nails. A mechanism for indoor fungal-mediated corrosion (biodeterioration) and biomineralisation of iron is proposed. The mechanism includes the establishment of a biofilm dominated by fungi, a corrosive electrolyte produced by fungal activities, transport of ions and biomineralisation of iron-rich minerals.

Culture-dependent and culture-independent methods were used to evaluate the microbial diversity in two hot springs of the Aljouf region in Saudi Arabia, including Qasr Kaff and Ain Hawas. Physicochemical characteristics of the springs were examined to establish their effect on the biodiversity of thermophilic bacteria and fungi. We employed culture-dependent techniques to study microbial diversity using four different complex media for bacteria and fungi. In addition, the direct count for algal populations from two springs was investigated. We surveyed the microbial diversity in water and sediment samples from both springs by denaturing gradient gel electrophoresis (DGGE) and clone library construction. Bacillariophycaea (18 species) was the most diverse group, followed by Cyanophyceaea. Bacterial isolates closer to the genera Bacillus spp., Geobacillus, Thermoactinomyces, and unidentified actinobacteria were recovered. Fungal isolates were related to Aspergillus, Pezizaceae, Penicillium, Acremoni...

Different protocols based on Illumina high-throughput DNA sequencing and denaturing gradient gel electrophoresis (DGGE)-cloning were developed and applied for investigating hot spring related samples. The study was focused on three target genes: archaeal and bacterial 16S rRNA and mcrA of methanogenic microflora. Shorter read lengths of the currently most popular technology of sequencing by Illumina do not allow analysis of the complete 16S rRNA region, or of longer gene fragments, as was the case of Sanger sequencing. Here, we demonstrate that there is no need for special indexed or tailed primer sets dedicated to short variable regions of 16S rRNA since the presented approach allows the analysis of complete bacterial 16S rRNA amplicons (V1-V9) and longer archaeal 16S rRNA and mcrA sequences. Sample augmented with transposon is represented by a set of approximately 300 bp long fragments that can be easily sequenced by Illumina MiSeq. Furthermore, a low proportion of chimeric sequences was observed. DGGE-cloning based strategies were performed combining semi-nested PCR, DGGE and clone library construction. Comparing both investigation methods, a certain degree of complementarity was observed confirming that the DGGE-cloning approach is not obsolete. Novel protocols were created for several types of laboratories, utilizing the traditional DGGE technique or using the most modern Illumina sequencing.

There has been much more damage to forests in the Slovak Republic in the second half of the 20th century than to other European countries. Forested mountain massifs have become a filter of industrial and transportation emissions from abroad, as well as from domestic origins. There are not only acidic deposits of sulphur and heavy metals present in forest soils, but other additional environmental problems, such as climate change, storms, fires, floods, droughts, are worsening the situation. Therefore, forest terrestrial ecosystems are becoming more vulnerable due to changes in natural and environmental conditions. In the High Tatra Mountains in Slovakia, which are protected as a national park, four internationally monitored localities were established after the windthrow disaster in 2004 and fire in 2005: REF, with intact forest, EXT, with extracted wood mass, NEX, with non-extracted wood mass, and FIR, the burnt locality. Soils from these localities were microbiologically analysed with special attention to fungi. Bacterial microbiota detected by high-throughput sequencing showed the prevalence of the genera Acidothermus, Mycobacterium, and Nocardia, and a very low presence of the genera Acidibacter, Burkholderia-Paraburkholderia, Optitus and the uncultured genus Desulfurellaceae H16 in the soil sample from the burnt locality when compared with the unburned sites. Additionally, soil mycocoenoses showed a low similarity between the locality with an intact forest ecosystem and the localities with extracted (REF&ndash, EXT) and non-extracted (REF&ndash, NEX) wood mass. There was no similarity with the burnt locality (FIR), where heat-resistant fungi dominated. It was shown that the windthrow disaster and subsequent extraction or non-extraction of wood mass did not affect the soil microbial communities or their development. On the other hand, the influence of fire was significant.

The microbial communities responsible for the degradation of poly(lactic acid)/poly(3-hydroxybutyrate) (PLA/PHB) blend foils were investigated in 1 year long laboratory soil burial experiments. Different PLA/PHB foils were tested: (a) PLA/PHB original transparent foil, (b) PLA/PHB carbon black filled foil and (c) PLA/PHB black foil previously exposed for 90 days to sun light. The microbiome diversity of these three types of foil was compared with that identified from soil/perlite sample at the beginning of experiment and that developed on a cellulose mat. Culture-dependent and culture-independent (DGGE-cloning) approaches together with PLA, PHB and PLA/PHB degradation plate assays were employed. The cultivation strategy combined with degradation tests permitted the isolation and evaluation of several PLA/PHB blend degrading microorganisms such as members of the genera Bacillus, Paenibacillus, Streptomyces, Rhodococcus, Saccharothrix, Arthrobacter, Aureobasidium, Mortierella, Absidia, Actinomucor, Bjerkandera, Fusarium, Trichoderma and Penicillium. The DGGE-cloning investigation increased the information about the microbial communities occurring during bioplastic degradation detecting several bacterial and fungal taxa and some of them (members of the orders Anaerolineales, Selenomonadales, Thelephorales and of the genera Pseudogymnoascus and Pseudeurotium) were revealed here for the first time. This survey showed the microbiome colonizing PLA/PHB blend foils and permitted the isolation of several microorganisms able to degrade the tested polymeric blends.

This microbiological survey was performed to determine the conservation state of a mummy in the Slovak castle of Krásna Hôrka and its surrounding environment. Culture-dependent identification was coupled with biodegradation assays on keratin, gelatin and cellulose. Next Generation Sequencing (NGS) using Illumina platform was used for a deeper microbial investigation. Three environmental samples were collected: from the glass of the sarcophagus, from the air inside it, and from the air of the chapel where the mummy is located. Seven different samples were taken from mummy's surface: from the left ear, left-hand palm, left-hand nail, left instep, right hand, abdomen and mineral crystals embedded within the skin. Three internal organ samples, from the lung, pleura and stomach, were also included in this study. Together, the culture-dependent and culture-independent analyses revealed that the bacterial communities present had fewer taxa than the fungal ones. The mycobiome showed the largest variability and included Epicoccum nigrum, Penicillium spp., Alternaria spp., Aspergillus spp., Cladosporium spp. and Aureobasidium pullulans; many other Ascomycota and Basidiomycota genera were detected by NGS. The most interesting results came from the skin mineral crystals and the internal organs. The hydrolytic assays revealed those microorganisms which might be considered dangerous 'mummy pathogens'. © 2018 Society for Applied Microbiology and John WileySons Ltd.

Microscopic fungi were isolated from different materials including muscles, bones, skin and funeral clothes from the mummified human remains of three members of the Kuffner's family and from the surrounding air environments. Their hydrolytic abilities such as cellulolytic, lipolytic, and proteolytic-keratinolytic were also assessed. The most isolated fungi, from human remains, belonged mainly to the species of Aspergillus (Aspergillus candidus, Aspergillus calidoustus, Aspergillus fumigatus, Aspergillus niger, Aspergillus sydowii, Aspergillus terreus, Aspergillus ustus, Aspergillus venenatus, Aspergillus versicolor, Aspergillus westerdijkiae) and Penicillium (Penicillium chrysogenum, Penicillium commune, Penicillium crustosum, Penicillium griseofulvum, Penicillium hordei, Penicillium polonicum). Aspergilli and penicillia were the predominant actors also in the air samples, but also many strains belonging to the Rhizopus group were isolated as well. Several fungi exhibited different hydrolytic ability, the most active isolated from human remains belonged to the species A. candidus, A. westerdijkiae, Coprinellus xanthothrix, P. chrysogenum, P. commune, P. griseofulvum and Scopulariopsis brevicaulis. The species recovered from the air displayed stronger deterioration characteristics as compared to human samples. This study can be considered one of few investigations focused on mummified human remains conserved in this kind of the environment.

Summary Roman Catacombs are affected by different kinds of biofilms that were extensively investigated in the last 14 years. In particular, the areas far from the lamps are often covered by white biofilms of different extension, consistency and nature. The aim of this paper is to describe the profile of the microbial community present in two areas of the Ocean's Cubiculum (CSC13), characterized by similar alterations described as white biofilms, by using a multistep approach that included direct microscopy observations, culture-dependent and culture-independent methodologies through the extraction of DNA and RNA directly from the sampled areas. In addition to this, we extracted the DNA directly from the Petri dishes containing R2A and B4 media after incubation and growth of bacteria. Our results evidenced that a complex bacterial community (mainly constituted by filamentous Actinobacteria, as well as Firmicutes and Proteobacteria) colonizes the two different white biofilms, and its detection, quantitative and qualitative, could be revealed only by different approaches, each method giving different information that only partially overlap.

Objectives Urban wastewater contains various micropollutants and a high number of different micro-organisms. Some bacteria in wastewater can attach to surfaces and form biofilm, which gives bacteria an advantage in the fight against environmental stresses. This work focused on analysis of bacterial communities in biofilms isolated from influent and effluent sewerage of a wastewater treatment plant (WWTP) in Bratislava, Slovakia. Methods Detection of biofilm microbiota was performed by culture-independent and -dependent approaches. The composition of bacterial strains was detected by denaturing gradient gel electrophoresis fingerprinting coupled with construction of 16S rRNA clone libraries. Analysis of the concentration of antibiotics and the prevalence of antibiotic-resistant coliforms, Enterococcus spp. and Staphylococcus spp. in sewerage was also studied. Results Biofilm collected at the inlet point was characterised primarily by the presence of Pseudomonas spp., Acinetobacter spp. and Janthinobacterium spp. clones, whilst members of the genus Pseudomonas were largely detected in biofilm isolated in outflow of the WWTP. Predominant antibiotics such as azithromycin, clarithromycin and ciprofloxacin were found in influent wastewater. The removal efficiency of these antibiotics, notably azithromycin and clarithromycin, was 30% in most cases. Conclusion The highest number of antibiotic-resistant bacteria, with a predominance of coliforms, was detected in samples of effluent biofilm. Multidrug-resistant strains in effluent biofilm showed very good biofilm-forming ability.

During the 20th century, synthetic polymers were greatly used in the field of art. In particular, the epoxy resins were used for both conservation and for creating sculptures. The biodeterioration of these polymers has not been adequately studied. The aim of this investigation was to examine the microflora responsible for the deterioration of an epoxy statue exposed to outdoor conditions. Fungal and bacterial microflora were isolated from the art object, clustered by fluorescence-ITS (internal transcribed spacer), identified by ITS and 16S rRNA sequencing and tested for their lipolytic abilities by three agar assays. Different algal, bacterial, cyanobacterial and fungal clone libraries were constructed. The surrounding airborne microflora was analyzed using culture-dependent and culture-independent approaches. The results indicated the presence, on the statue surface, of an interesting and differentiate microbial community composed of rock-inhabiting members, algal photobionts (Trebouxia spp., Chloroidium ellipsoideum and Chlorella angustoellipsoidea), Cyanobacteria (Leptolyngbya sp., Phormidium sp., Cylindrospermum stagnale, Hassallia byssoidea and Geitlerinema sp.), black yeasts related to the species Friedmanniomyces endolithicus, Pseudotaeniolina globosa, Phaeococcomyces catenatus and Catenulostroma germanicum and several plant-associated fungi. This investigation provides new information on the potential microfloral inhabitants of epoxy resin discovering a new ecological niche, occupied mainly by several members of rock-colonizing microbial species.

The aim of this study was to investigate the yeast and fungal microflora diversity in wine-related samples of the cultivars Veltlin zelený (Gruner Veltliner) and Frankovka modra (Blaufrankisch) produced in the Small Carpathian region (Slovakia) using different eukaryotic identification approaches. For phenotypic identification we performed auxotrophy profiling using the API 20C AUX kit (bioMerieux, Marcy l'Etoile, France) and culture on selective lysine medium. Genotypic identification was done by real-time PCR assays specific for different wine yeasts and by sequencing of the internal transcribed spaced (ITS) region (ITS1–5.8S rRNA–ITS2 region). An effective culture-independent approach was also employed in the form of analysis of the ITS2 region using fluorescence–ITS PCR (f-ITS PCR), cloning and sequencing. The ITS sequencing results were concordant with the results of the selective culture and real-time PCR assay, but for several strains they did not agree with the results of auxotrophy profiling. In the culture-independent approach, sequences of clones included 100 % matches to Hanseniaspora uvarum, Candida zemplinina, Metschnikowia pulcherrima, Aureobasidium pullulans, Saccharomyces cerevisiae and Issatchenkia terricola, as well as two novel sequences with a 5-bp difference to Sporobolomyces marcillae. The data from this study, and particularly the data from the culture-independent investigation, contribute to the picture of the wine microbial community of two cultivars important in wine production in Slovakia and Central Europe.

The use of indigenous bacterial strains is a valuable bioremediation strategy for cleaning the environment from hydrocarbon pollutants. The isolation and selection of hydrocarbon-degrading bacteria is therefore crucial for obtaining the most promising strains for site decontamination. Two different media, a minimal medium supplemented with a mixture of polycyclic aromatic hydrocarbons and a MS medium supplemented with triphenyltetrazolium chloride, were used for the isolation of bacterial strains from two hydrocarbon contaminated soils and from their enrichment phases. The hydrocarbon degradation abilities of these bacterial isolates were easily and rapidly assessed using the 2,6-dichlorophenol indophenol assay. The diversity of the bacterial communities isolated from these two soil samples and from their enrichment phases was evaluated by the combination of a bacterial clustering method, fluorescence ITS-PCR, and bacterial identification by 16S rRNA sequencing. Different PCR-based assays were performed in order to detect the genes responsible for hydrocarbon degradation. The best hydrocarbon-degrading bacteria, including Arthrobacter sp., Enterobacter sp., Sphingomonas sp., Pseudomonas koreensis, Pseudomonas putida and Pseudomonas plecoglossicida, were isolated directly from the soil samples on minimal medium. The nahAc gene was detected only in 13 Gram-negative isolates and the sequences of nahAc-like genes were obtained from Enterobacter, Stenotrophomonas, Pseudomonas brenneri, Pseudomonas entomophila and P. koreensis strains. The combination of isolation on minimal medium with the 2,6-dichlorophenol indophenol assay was effective in selecting different hydrocarbon-degrading strains from 353 isolates.

Studied technosols represent a unique system of a 50-year-old environmental burden after dam failure of coal-ash pond. The released ashes rich in arsenic with a thickness of 1–2 m were covered by a 40-cm thick layer of soil. Long-term exposure and selection pressure of elevated concentrations of arsenic (a range of 93–634 μg/g) induced the formation of the specific adapted autochthonous microorganisms. The phylum Proteobacteria was identified as a dominant phylum in the soils and represented only by one class—Gammaproteobacteria with six species. The species of phylum Firmicutes, Bacteroidetes and Actinobacteria were also identified. Thirty-three species of identified autochthonous microscopic fungi belong to 18 genera with the most abundant Mortierella alpina (Zygomycota). The most frequent identified mycobiota belongs to genera Penicillium, Aspergillus, Trichoderma and Alternaria. The isolates of Alternaria triticina, Bionectria ochroleuca, Chrysosporium queenslandicum, Exophiala psychrophila, Metarhizium robertsii, Trichoderma rossicum and Phlebia acerina were identified for the first time in Slovakia. Despite the stimulation of autochthonous community by nutrient medium and augmentation by native species, As leachability was relatively low—on average 5.63 wt.%, 9.23 wt.% and 17.04 wt.% of the total As for inoculated Pseudomonas chlororaphis ZK-1, Pseudomonas putida ZK-5 and Aspergillus niger, respectively. The highest As leachability was achieved through biostimulation of autochthonous microbiota using liquid SAB medium (34.73 wt.% of total As content). Additionally, microbial activity was efficient in the biovolatilization of As from soils (∼70 wt.% of the total As volatilized). It appears that bioremediation using microorganisms represents one of the possible ways of As removal from soils containing coal-combustion ashes with elevated concentrations of As.

The gelatin-silver halide black and white prints represent an enormous photography heritage with a great value. Unaesthetic phenomena, the foxing stains that are caused by microbial growth on surface, have been described in stamps, drawings, books, and tissues but, until now, scarcely for photographic materials. In this study, a combination of various techniques, including culture-dependent and culture-independent approaches (RNA and DNA analysis), scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS) and μ-Raman spectroscopy supported by X-ray fluorescence analysis (XRF), permitted to describe the microbial contamination dynamics of foxing stains present on the surface of two gelatin-silver halide photographs. The investigation provided also information on the effects of microbial activity on the materials’ chemistry of the two prints. The action of microbial community resulted locally in either (a) formation of mixed aluminum-iron-potassium phosphate compounds that could be attributed to the hydrolytic activity of bacteria, (b) leaching of barite, (c) precipitation of a mixture of oxides, and (d) a change in the barium sulfate chemical structures.

Different types of biofilms are widespread on lithic faces of the Catacombs of Domitilla (Rome, Italy) due to the favorable microclimatic conditions (temperature, high RH% and low irradiance). The biofilm, once established, becomes particularly dangerous due to the coverage of valuable surfaces causing spoilage, softening of materials and mineral precipitation. It is common practice to treat these surfaces with biocides in order to eradicate the microorganisms present. The aim of the present research was to compare the changes occurring to the microbial community present in the biofilm in one site of the Catacombs of Domitilla (CD15) before and after a biocide treatment (a mixture of quaternary ammonium compounds and octylisothiazolone, OIT), applied for a one month period. A multistep approach was followed, based on microscopy, cultural methods and molecular techniques (f-ITS and 16S rDNA sequencing), for the phenotypic and genetic analysis of the culturable microbial population. Our results highlighted that the biocide treatments had little effect against cyanobacteria, while the bacterial population increased in numbers but changed drastically in terms of diversity. In fact, some bacteria proliferate at the expense of the organic matter released by dead microorganisms as demonstrated by laboratory tests. Further, our data describe how the microbial interaction can have different responses depending on the favorable conditions for one kind of microorganism in respect to the others. This study exemplifies the real risks of applying biocide treatments on complex microbial communities and pinpoints the necessity of subjecting treatments to monitoring and reassessment. Moreover, the work showed the potential of bacteria isolated after the treatment for use, under controlled conditions, in combatting unwanted microbial growth in that they possess a positive tropism toward stressed microorganisms and high hydrolytic enzymatic activity against cell components ( e . g . cellulose, chitin and pectin). A tentative protocol is proposed.

A multistep procedure was applied in order to investigate the yeasts diversity during the must fermentation of two wine varieties Frankovka Modra (Blaufrankisch) and Veltlinske Zelene (Gruner Veltliner) from two different Slovakian wine-producing areas. The procedure combined a cultivation step with a PCR-based selection step (f-ITS PCR; fluorescence internal transcribed spacer 2 polymerase chain reaction), and a final identification step oriented to the sequencing of ITS1-5.8S rDNA-ITS2 region of the selected yeasts. The strategy demonstrated to be suitable for the rapid analysis of a large panel of isolates. The f-ITS PCR was able to discriminate the isolated yeasts by one PCR run and capillary electrophoresis separation. The investigation showed the presence of non-Saccharomyces strains on grapes and during the initial and middle stages of must fermentation. Members of the genus Saccharomyces appeared only during the middle fermentation stage and they were the predominant species until the final phase of the process. The presented simple and reliable multistep procedure can be useful for the investigation of the yeast microflora from any wine-producing area and also for studying other kinds of food environments.

The microbial diversity of different kinds of stains present on the surface of 14 historical documents (nine parchments and five paper letters) was evaluated through a combination of cultural and molecular methods. The samples were recovered using adhesive tape and swabs and were afterwards treated in two different ways: (1) direct inoculation on agar plates; or (2) suspension in physiological solution and plating in specific plates for the growth of bacteria and fungi. The isolated microorganisms, before identification, were selected by two different PCR-based methods – f-ITS and f-CBH, for bacteria and fungi, respectively. The f-ITS method is based on the amplification of the internal transcribed sequence between the bacterial 16S and 23S rDNA. The f-CBH method is a new molecular selection tool oriented to the fungal cellobiohydrolase gene. Both PCR selection methods produced typical profiles, which clustered the isolates in order to reduce them for subsequent sequencing identification through the amplification of the fungal 28S rRNA and the bacterial 16S rRNA genes. The cellulolytic and proteolytic abilities were screened through the use of three plate assays, the Ostazin Brilliant Red H-3B (OBR-HEC), milk agar, and gelatin agar. Massilia timonae , Lysobacter dokdonensis , and strains belonging to the genus Bacillus sp., Microbacterium sp., and Curtobacterium sp. with different fungal members such as Aspergillus fumigatus , Penicillium commune , Mucor spinosus , and Phoma herbarum (all recovered from paper) displayed both biodegradative activities. The parchment isolates with a marked proteolytic activity were Bacillus cereus , Staphylococcus epidermidis , Pseudomonas oryzihabitans , Virgibacillus sp., Micromonospora sp., and again members of the fungal genera Penicillium , Mucor , and Phoma .

Aims: To develop a novel PCR-based method able to detect potential cellulolytic filamentous fungi and to classify them exploiting the amplification of the cellobiohydrolase gene (cbh-I) and its polymorphism. Methods and Results: A mixed approach including the combination of (i) fungal cultivation and isolation, (ii) classification of fungal isolates through the amplification of the cbh gene using a fluorescently labelled primer (f-CBH-PCR) and (iii) final fungal identification based on amplification and sequencing of the ITS1-5.8S rDNA-ITS2 region of the selected fungal strains was developed. By this approach, it was possible to screen 77 fungal strains belonging to 14 genera and 26 species. Conclusions: The f-CBH-PCR permitted the discrimination of fungal species, producing typical f-CBH profiles. Significance and Impact of the Study: In this study, the cbh gene was used as a preliminary classification tool able to differentiate among themselves the fungal members isolated from indoor museum items and surrounding environment. Such mixed approach consented the fast identification of all isolated fungal strains. The f-CBH-PCR method demonstrated its discrimination power, and it can be considered as a new molecular system suitable for the classification of fungal strains isolated from different environments.

Aims: The investigation of yeast microflora during the must fermentation of two wine varieties (Frankovka modra – Blaufrankisch and Veltlinske zelene – Gruner Veltliner) from two consecutive vintages was performed using a three-step approach. Methods and Results: The investigation strategy consisted of the combination of yeast cultivation, selection of the isolated yeasts based on the amplification of internal transcribed spacer 2 using a fluorescence-labelled primer (f-ITS-PCR) and a final identification step based on amplification and sequencing of the ITS1-5.8S rDNA-ITS2 region of the selected yeasts. By this three-step approach, it was possible to screen 433 yeasts isolates that belonged to 13 different species. Conclusions: The f-ITS-PCR allowed the unambiguous differentiation of all isolated yeast species that produced their typical f-ITS-PCR profile. Significance and Impact of the Study: This is one of few reports that treat the yeast diversity in Slovakian wines and in two varieties largely cultivated in Central Europe. The three-step approach permitted the rapid and reliable identification of isolated yeasts. The f-ITS-PCR with its good discrimination power can represent a suitable molecular tool for the selection of yeast members recovered from food or other environments.

Since the yeast flora of Slovakian enology has not previously been investigated by culture-independent methods, this approach was applied to two most common cultivars Frankovka (red wine) and Veltlin (white wine), and complemented by cultivation. Model samples included grapes, initial must, middle fermenting must and must in the end-fermentation phase. The cultured isolates were characterized by length polymorphism of rDNA spacer two region using fluorescence PCR and capillary electrophoresis (f-ITS PCR), and some were identified by sequencing. The microbial DNA extracted directly from the samples without cultivation was analysed by f-ITS PCR, amplicons were cloned and sequenced. The use of universal fungal primers led to detection of both yeasts and filamentous fungi. The amplicon of highest intensity and present in all the samples corresponded to Hanseniaspora uvarum. Other species demonstrated by both approaches included Saccharomyces sp., Metschnikowia pulcherrima or M. chrysoperlae, Candida zemplinina, Cladosporium cladosporioides, Botryotinia fuckeliana, Pichia anomala, Candida railenensis, Cryptococcus magnus, Metschnikowia viticola or Candida kofuensis, Pichia kluyveri or Pichia fermentas, Pichia membranifaciens, Aureobasidium pullulans, Alternaria alternata, Erysiphe necator, Rhodotorula glutinis, Issatchenkia terricola and Debaryomyces hansenii. Endemism of Slovakian enological yeasts was suggested on the level of minor genetic variations of the known species and probably not accounting for novel species. The prevalence of H. uvarum over Saccharomyces sp. in the samples was indicated. This is the first culture-independent study of Slovakian enology and the first time f-ITS PCR profiling was used on wine-related microbial communities.

This study is one of the few investigations which analyze albumen prints, perhaps the most important photographic heritage of the late 19th and early 20th centuries. The chemical composition of photographic samples was assessed using Fourier-transform infrared spectroscopy and X-ray fluorescence. These two non-invasive techniques revealed the complex nature of albumen prints, which are composed of a mixture of proteins, cellulose and salts. Microbial sampling was performed using cellulose nitrate membranes which also permitted the trapped microflora to be observed with a scanning electron microscope. Microbial analysis was performed using the combination of culture-dependent (cultivation in different media, including one 3% NaCl) and culture-independent (bacterial and fungal cloning and sequencing) approaches. The isolated microorganisms were screened for their lipolytic, proteolytic, cellulolytic, catalase and peroxidase activities. The combination of the culture-dependent and -independent techniques together with enzymatic assays revealed a substantial microbial diversity with several deteriogen microorganisms from the genera Bacillus, Kocuria, Streptomyces and Geobacillus and the fungal strains Acrostalagmus luteoalbus, Bjerkandera adusta, Pleurotus pulmonarius and Trichothecium roseum.

Studying the culturable portion of environmental bacterial populations is valuable for understanding the ecology, for discovering taxonomically interesting isolates and for exploiting their enzymatic abilities. In this study, diverse water-related samples, iced water (3 °C) from river, the sediment (29 °C) and water (55 °C) of a hot-spring, were investigated by two cultivation strategies, Dry and novel Wet approach. The isolates were clustered by fluorescent internal transcribed spacer PCR and identified by 16S rRNA sequencing. Several bacterial groups were also sub-typed through the application of Random Amplified Microsatellite Polymorphisms method. A broad enzymatic screening of all bacterial isolates was performed in order to assess the proteolytic, cellulolytic, lipolytic, esterolytic, amylolytic properties, as well as catalase and peroxidase activities. The Wet cultivation demonstrated to be suitable for the isolation of potential new species belonging to genera Massilia, Algoriphagus, Rheinheimera and Pandoraea. Valuable microbial resources with extensive enzymatic activities were recognized among the psychrophilic (Pantoea brenneri and Serratia sp.), mesophilic (Pandoraea, Massilia, Pseudomonas, Stenotrophomonas, Bacillus and Aeromonas) and thermophilic bacteria (Aeribacullus pallidus and Geobacillus kaustophilus). The experimental strategy developed in this study includes simple investigation tools able to reveal the genetic and enzymatic peculiarities of isolated microorganisms. It can be applied to different kinds of aquatic samples and extreme environments similar to those described in this study.

This is the first study focused to bacterial diversity and dynamic during the vinification of two important Central Europe grape vines: Blaufrankisch and Gruner Veltliner. The investigation strategy included culture-dependent and culture-independent approaches. Four different agar media were utilized for the isolation of various bacteria occurring in several fermentation stages. The isolates were clustered by fluorescent-ITS PCR and, one or more representatives of each cluster, were identified by 16 rRNA gene sequencing. The culture-independent approach, based on 16S rRNA gene amplification, combined the denaturing gradient gel electrophoresis (DGGE) method and the construction of bacterial clone library for each wine fermentation step. A complex bacterial community was identified, comprising different lactic acid bacteria and acetic acid bacteria, such as Leuconostoc spp., Lactobacillus spp. and Gluconobacter spp. Other OTUs and bacterial isolates embraced the Actinobacteria, Bacilli, Alpha-, Beta- and Gamma-proteobacteria classes. Different taxa already detected by recent studies, such as Sphingomonas, Variovorax, Pantoea, Enterobacter and Tatumella, were detected confirming the continuous occurrence of these kinds of bacteria in wine environment. Moreover, novel genera (Amycolatopsis, Hydrogenophilus, Snodgrassella, Telluria, Gilliamella, Lelliottia, and Lonsdale quercina) never detected before were recognized, too. The role of these, until now anonymous, bacteria during vinification deserves investigation, which could open a new research field in wine technology.

Three innovative PCR-based methods (fluorescent-ITS, fluorescent-CBH and ITS-PCR DGGE) were tested using a reference set of nine strains of Scedosporium from the CBS fungal collection. Cellulolytic, lipolytic and proteolytic potential and the ability to dissolve CaCO3 of the strains were evaluated in vitro by means of agar assays. f-ITS profiles almost recognized main species, although included "Pseudallescheria" ellipsoidea and the Scedosporium boydii CBS 117432 and CBS 120157 in the same cluster. All strains successfully produced DNA polymorphisms by f-CBH amplification which divided them into three different groups. The DGGE approach separated the strains studied into other five clusters which in some case were not matching with species. Strains tested were monomorphic in possessing strong proteolytic and lipolytic activities. The comparison of the three PCR-based genotyping approaches, together with biodegradation ability screening, displayed an intraspecies variability in S. boydii, interfering with unambiguous species delimitation.

During the 20th century, synthetic polymers were greatly used in the field of art. In particular, the epoxy resins were used for both conservation and for creating sculptures. The biodeterioration of these polymers has not been adequately studied. The aim of this investigation was to examine the microflora responsible for the deterioration of an epoxy statue exposed to outdoor conditions. Fungal and bacterial microflora were isolated from the art object, clustered by fluorescence-ITS (internal transcribed spacer), identified by ITS and 16S rRNA sequencing and tested for their lipolytic abilities by three agar assays. Different algal, bacterial, cyanobacterial and fungal clone libraries were constructed. The surrounding airborne microflora was analyzed using culture-dependent and culture-independent approaches. The results indicated the presence, on the statue surface, of an interesting and differentiate microbial community composed of rock-inhabiting members, algal photobionts (Trebouxia spp., Chloroidium ellipsoideum and Chlorella angustoellipsoidea), Cyanobacteria (Leptolyngbya sp., Phormidium sp., Cylindrospermum stagnale, Hassallia byssoidea and Geitlerinema sp.), black yeasts related to the species Friedmanniomyces endolithicus, Pseudotaeniolina globosa, Phaeococcomyces catenatus and Catenulostroma germanicum and several plant-associated fungi. This investigation provides new information on the potential microfloral inhabitants of epoxy resin discovering a new ecological niche, occupied mainly by several members of rock-colonizing microbial species.