This study focuses on the magnificent decoration of a painted and gilded wooden panel with signs of fungal biodeterioration caused by Aspergillus species in the Mausoleum of Sultan al-Ashraf Qaytbay, Cairo, Egypt. Numerous spectroscopic analyses and investigation techniques, including Scanning Electron Microscope Equipped with Energy Dispersive X-ray analysis (SEM–EDX), Fourier Transform Infrared analysis (FTIR), and X-Ray Diffraction (XRD) have been used to study the materials that comprise this painted and gilded wooden panel composition. Aspergillus niger, A. flavus, and A. terreus were recognized as isolated fungi, and their accession numbers are OQ820164, OQ820163, and OQ820160, respectively. The findings showed that the wooden support is of pinewood (Pinus halepensis), the white priming layer on top of the wooden support was identified as gypsum, the blue paint layer has been proposed to be Azurite, Au (gold) was the primary composition of the gilding layer, while Pb (lead) was detected in some spots, suggesting the use an alloy of gold with lead, and finally, animal glue was the bonding medium. Based on these findings, mimic samples with identical substrates and structural components have been designed, and the biodeterioration signs by the growing of the three Aspergillus species—A. niger, A. flavus and A. terreus were evaluated via SEM and color change. However, A. niger was discovered with density growth on surfaces of pinewood, gypsum, and Azurite and with less growth on the gilding layer after 6-month incubation. This contrasts with A. terreus and A. flavus, which had greater density growth on Azurite and stucco than on pinewood and less growth on the gilding layer. The used analytical methods with detailed analyses revealed the novelty and significant future aspects of the conservation of the painted and gilded wooden panel. Particularly given that this location is used for prayer and is crowded with people five times a day, which increases the accumulation of fungi and negatively affects both the historic Mosque and the worshippers' health.
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