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Palm leaf manuscripts were a prevalent literary medium from South Asia and Southeast Asia prior to the widespread use of paper. This study focuses on the analysis of historical palm leaf manuscripts from South and Southeast Asia. Sample palm leaf manuscripts from Burma were used as a case study; simulated palm leaf manuscripts were also created as a reference for comparison. The anatomy, chemical composition, and mechanical properties of the manuscripts were analyzed to find various forms of deterioration, including damage, fractures, pollution, acidification, and microbial deterioration. Specifically, the S₁–S₃ layers of the cell walls exhibited complete cracking, and the S₂ layer showed numerous circular or nearly circular cavities caused by microbial erosion, while the middle lamella remained intact. The severe degradation of polysaccharides and pectin, accompanied by an increase in the relative content of lignin, caused the historical manuscripts to become more brittle. Additionally, the tensile strengths of historical palm leaf manuscripts were markedly reduced; their longitudinal tensile strength was significantly greater than their transverse tensile strength. This study can contribute to a better understanding of the deterioration process of historical palm leaf manuscripts and provide valuable insights for their restoration and preservation.