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Alcobacenses are one of the most important Portuguese collections of Monastic codices. During a campaign study to further advance our knowledge on this important legacy and its state of conservation, a heavily deteriorated codex (ALC.338) stood out due to the presence of purple stains on the lower margins on virtually all folios. In some of them, the purple stains are accompanied by yellow and brown patches as well. Also noticed was the presence of these purple spots on the white tawed leather covering the codex. The codex was analyzed macro and microscopically, chemically and biologically using state-of-the-art technology such as next-generation sequencing. The biological deterioration associated with the appearance of these purple spots arises from the inside out and reveals significant abundance of the bacterial genus Saccharopolyspora sp., which accounted for 86% of the bacterial genera identified. For the fungal population, the most common genus was Malassezia sp. Visual inspection of the sample confirmed the presence of fungal and bacterial biofilms. Pressing questions, comparison to similar studies and future perspectives are also given.<\/jats:p>","DOI":"10.3390\/heritage5040212","type":"journal-article","created":{"date-parts":[[2022,12,12]],"date-time":"2022-12-12T05:42:00Z","timestamp":1670823720000},"page":"4100-4113","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Underneath the Purple Stain"],"prefix":"10.3390","volume":"5","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7194-5496","authenticated-orcid":false,"given":"Catarina","family":"Pinheiro","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio HERCULES, Heran\u00e7a Cultural, Estudos e Salvaguarda, Largo Marqu\u00eas de Marialva, 8, 7000-809 \u00c9vora, Portugal"}]},{"given":"Ana Z\u00e9lia","family":"Miller","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio HERCULES, Heran\u00e7a Cultural, Estudos e Salvaguarda, Largo Marqu\u00eas de Marialva, 8, 7000-809 \u00c9vora, Portugal"},{"name":"Instituto de Recursos Naturales y Agrobiolog\u00eda de Sevilla (IRNAS-CSIC), Av. Reina Mercedes 10, 41012 Sevilla, Spain"}]},{"given":"Patr\u00edcia","family":"Vaz","sequence":"additional","affiliation":[{"name":"Departamento de Conserva\u00e7\u00e3o e Restauro, Faculdade de Ci\u00eancias e Tecnologia, Universidade Nova de Lisboa, Campus Caparica, 2829-516 Caparica, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5409-6990","authenticated-orcid":false,"given":"Ana Teresa","family":"Caldeira","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio HERCULES, Heran\u00e7a Cultural, Estudos e Salvaguarda, Largo Marqu\u00eas de Marialva, 8, 7000-809 \u00c9vora, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9889-381X","authenticated-orcid":false,"given":"Concei\u00e7\u00e3o","family":"Casanova","sequence":"additional","affiliation":[{"name":"Departamento de Conserva\u00e7\u00e3o e Restauro, Faculdade de Ci\u00eancias e Tecnologia, Universidade Nova de Lisboa, Campus Caparica, 2829-516 Caparica, Portugal"},{"name":"LAQV\u2014REQUIMTE, Universidade Nova de Lisboa, Campus Caparica, 2829-516 Caparica, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2022,12,11]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1080\/17546559.2016.1221116","article-title":"Beatus manuscripts under the microscope: The Alcoba\u00e7a Beatus and the Iberian Cistercian tradition revisited","volume":"8","author":"Melo","year":"2016","journal-title":"J. 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