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Abstract Potential high bio-deterioration of certain microorganisms due to their metabolic activities on organic materials causes serious problems in the conservation of cultural heritage. This study aims to isolation and identification of causative biodeterioration microbes and create a new approach for the treatment of biodeteriorated paper manuscripts. Macroscopic observations of collected manuscripts, isolation of microbes from the historical samples, microscopic identification for fungal isolates,identification of bacterial isolates using morphological, physiological and Vitek 2 methods were carried out in this study.Molecular identification showed thateleven bacterial species and fifteen fungal species were identified using 16S rRNA and ITS sequences, respectively. Paper models conservation effect of Ag-NPs, ZnO-NPs, AgNO3 and Zn(CH3CO2)2.2H2O against strains Bacillus subtilis, Aspergillus niger and Penicillium chrysogenum was evaluated. Color change (CIE L*a*b*), tensile strength and elongation, FT-IR and SEM were assessed to know the effects of microorganisms and materials used on paper properties.The manuscript under the title 2Al-Mosamara Sharh Al-Mosayara3 , dated back to 1124 A.H, and deposit in Al-Azhar Library, Cairo, was used for the applied study. Analytical techniques and investigations used for the evaluation of the manuscript state were optical microscope, investigationof black ink, acidity ratio measurement, FTIR, SEM, EDX,XRD, andmicrobiological investigation |