## 2 Materials and Methods
### Fungal Isolation and Identification  
Each nitrocellulose membrane used for sampling was washed with 2 mL of sterile 0.9% (\(w/v\)) NaCl solution, and the resulting suspension was collected. Per resulting sample, a volume of 100 \(\upmu\)L was inoculated in four different culture media: Potato Dextrose Agar (PDA; Difco, Sparks, MD, USA); Malt Extract Agar (MEA; Difco, Sparks, MD, USA); Dichloran-glycerol Agar (DG-18; Oxoid, Basingstoke, UK); and Blood Agar Base (BA, Sigma-Aldrich, St. Louis, MO, USA), in quintuplicate. Streptomycin was added to all media at a concentration of 0.5 gL\({}^{-1}\) to prevent bacterial growth. Inoculated media plates were incubated at 25 \(\pm\) 2 \({}^{\circ}\)C aerobically and left in the dark for 3 months. All emerging colonies in each culture medium with significantly different morphology were further isolated to axenic cultures onto both PDA and the original culture medium they were isolated from and incubated at 25 \(\pm\) 2 \({}^{\circ}\)C aerobically. When significant mycelium growth was observed,DNA extraction of the pure cultures was performed with the REDextract-N.AmpTM Plant PCR Kit (Sigma Aldrich, USA), with several modifications, as described by [PERSON] et al. [35]. DNA obtained was used for PCR amplification of the Internal Transcribed Spacer (ITS), RNA Polymerase II Second Largest Subunit (_Rpb2_), \(\beta\)-tubulin (_Hub_) and Calmodulin (_CaM_) regions, using the ITS1-F/ITS4 [36; 37], RPB2-5f/RPB2-7 cR [38; 39], Bt2a/Bt2b [40] and CMD5/CMD6 [41] primer pairs, respectively. ITS region amplification was performed for all isolates, while _Hub_ was only used for isolates identified as belonging to the _Aspergillus_, _Talaroumyces_ and _Penicillium_ genera, _Rpb2_ for those belonging to the _Trichoderma_ genera and _CaM_ for the _Aspergillus_ and _Talaroumyces_ isolates. PCR reactions, consisting of 12.5 \(\upmu\)L of NZYTaq Green Master Mix (NZYTech(tm), Lisboa, Portugal), 1 \(\upmu\)L of each primer (10 mM), 9.5 \(\upmu\)L of ultra-pure water and 1 \(\upmu\)L of template DNA, for a final amplification volume of 25 \(\upmu\)L, were performed using an ABI GeneAmpTM 9700 PCR System (Applied Biosystems, Waltham, MA, USA). The amplification conditions for ITS followed [PERSON] et al. [35], _Btub_ followed [PERSON] et al. [42] and for _Rpb2_ and _CaM_ the protocol was in accordance with [PERSON] and [PERSON] [43]. Confirmation of the amplification of the ITS, _Rpb2_, _Hub_ and _CaM_ regions was conducted through agarose gel electrophoresis (1.2%) stained with GreenSafe Premium (NZYTech(tm), Lisboa, Portugal) and visualized in a Molecular Imager Bio-Rad Gel Doc XR(tm) (Bio-Rad, Hercules, CA, USA). Obtained amplicons were purified and sequenced using an ABI 3730 xl DNA Analyzer system (96 capillary instruments) at STABVIDA, Portugal. Obtained DNA sequences were analyzed using the BioEdit Sequence Alignment Editor(r) v.7.2.5.0 \"[[https://bioedit.software.informer.com/download/](https://bioedit.software.informer.com/download/)]([https://bioedit.software.informer.com/download/](https://bioedit.software.informer.com/download/)) (accessed on 10 October 2024)\" and deposited in the GenBank database with the accession numbers PQ433809-PQ433851 and PQ439731-PQ439800. Figure 1: Sampled artifacts showing visible fungal contamination: (**a**) ANT.Moç490; (**b**) ANT.Moç349; (**c**) MCUC2013.1.3; (**d**) ANT.Moç344. Circled areas indicate sampled sites; in the case of ANT.Moç490, samples were divided based on the sampled material (wood and leather). Similarity searches were performed using the National Center for Biotechnology Information nucleotide database (NCBI's) online Basic Local Alignment Search Tool (BLAST), using the somewhat similar sequences (blastn) algorithm of BLAST [44].