{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,18]],"date-time":"2026-01-18T12:43:49Z","timestamp":1768740229865,"version":"3.49.0"},"reference-count":31,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2021,1,13]],"date-time":"2021-01-13T00:00:00Z","timestamp":1610496000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100010665","name":"H2020 Marie Sk\u0142odowska-Curie Actions","doi-asserted-by":"publisher","award":["766311"],"award-info":[{"award-number":["766311"]}],"id":[{"id":"10.13039\/100010665","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["CMD"],"abstract":"<jats:p>An experimental study was conducted to assess the nature and extent of the biodeterioration of the limestone in the Batalha Monastery in Portugal. Stone fragments covered with microbial biofilms and lichenous crusts were investigated using Optical Microscopy (OM), Low Vacuum Scanning Electron Microscopy with Energy Dispersive Spectroscopy (LV-SEM + EDS), and X-ray micro-Diffractometry (\u03bc-XRD). Microbial samples were collected from the stone surface, cultured, and analyzed with NGS metagenomic DNA test to classify the bacterial communities associated with the formation of the biofilms. Particulate air pollutants collected on Pall GN-6 paper filters using a cascade impactor were characterized by SEM-EDS + NGS. The results showed that lichens play a major role in biodeterioration by promoting both physical and chemical attack on the limestone substrate via hyphae mechanical penetration along calcite inter-crystalline spaces, the dissolution\/leaching of calcite minerals, and the precipitation of secondary minerals such as Ca-oxalates within the stone porosity framework. DNA analyses identified the bacterial communities within the biofilms and their relative abundances. Air quality monitoring results suggest that the microbial population colonizing the monastery limestone could at least partially be derived from the dry and wet deposition of airborne biological particles on the stone surfaces and that S, N, and P-rich air pollutants may have provided nutrients and energy for the bacteria communities, thus indirectly facilitating biofilm formation, the growth of a lichenous crusts, and limestone biodeterioration effects.<\/jats:p>","DOI":"10.3390\/cmd2010002","type":"journal-article","created":{"date-parts":[[2021,1,13]],"date-time":"2021-01-13T21:50:54Z","timestamp":1610574654000},"page":"31-45","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["Biodegradation and Microbial Contamination of Limestone Surfaces: An Experimental Study from Batalha Monastery, Portugal"],"prefix":"10.3390","volume":"2","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1983-6057","authenticated-orcid":false,"given":"Yufan","family":"Ding","sequence":"first","affiliation":[{"name":"Hercules Laboratory, University of \u00c9vora, 7000-812 \u00c9vora, Portugal"},{"name":"Department of Applied Science and Technology, Polytechnic of Turin, 10138 Torino, Italy"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1797-1404","authenticated-orcid":false,"given":"Catia Sofia Clemente","family":"Salvador","sequence":"additional","affiliation":[{"name":"Hercules Laboratory, University of \u00c9vora, 7000-812 \u00c9vora, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5409-6990","authenticated-orcid":false,"given":"Ana Teresa","family":"Caldeira","sequence":"additional","affiliation":[{"name":"Hercules Laboratory, University of \u00c9vora, 7000-812 \u00c9vora, Portugal"},{"name":"Department of Chemistry, University of \u00c9vora, 7000-812 \u00c9vora, Portugal"}]},{"given":"Emma","family":"Angelini","sequence":"additional","affiliation":[{"name":"Department of Applied Science and Technology, Polytechnic of Turin, 10138 Torino, Italy"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9744-0808","authenticated-orcid":false,"given":"Nick","family":"Schiavon","sequence":"additional","affiliation":[{"name":"Hercules Laboratory, University of \u00c9vora, 7000-812 \u00c9vora, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2021,1,13]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Ahmadjian, V. (1973). Resynthesis of lichens. The Lichens, Academic Press.","DOI":"10.1016\/B978-0-12-044950-7.50023-4"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1016\/S0964-8305(00)00109-8","article-title":"Biodeterioration of stone: A review","volume":"46","author":"Warscheid","year":"2000","journal-title":"Int. Biodeterior. Biodegrad."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1038\/ismej.2014.138","article-title":"Exploring functional contexts of symbiotic sustain within lichen-associated bacteria by comparative omics","volume":"9","author":"Grube","year":"2015","journal-title":"ISME J."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.ibiod.2019.05.008","article-title":"Fungal diversity and distribution across distinct biodeterioration phenomena in limestone walls of the old cathedral of Coimbra, UNESCO World Heritage Site","volume":"142","author":"Portugal","year":"2019","journal-title":"Int. Biodeterior. Biodegrad."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/j.culher.2018.07.008","article-title":"Limestone biodeterioration: A review on the Portuguese cultural heritage scenario","volume":"36","author":"Pinheiro","year":"2019","journal-title":"J. Cult. Herit."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"3476","DOI":"10.1099\/mic.0.032508-0","article-title":"Biodiversity of cyanobacteria and green algae on monuments in the Mediterranean Basin: An overview","volume":"155","author":"Macedo","year":"2009","journal-title":"Microbiology"},{"key":"ref_7","unstructured":"Prada, J.L., Pug\u00e9s, M., and Rocabayera, R. (2002). Estudio de las patologias del Retablo del Altar Mayor de la S\u00e9 da Guarda. Patrim\u00f3nio Estudos, Instituto Portugu\u00eas do Patrim\u00f3nio Arquitect\u00f3nico."},{"key":"ref_8","first-page":"627","article-title":"Endolithic microorganisms in the biodeterioration of the tower of Belem","volume":"4","author":"Ascaso","year":"1998","journal-title":"Int. Z. F\u00fcrBauinstandsetz."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0964-8305(01)00097-X","article-title":"In situ evaluation of the biodeteriorating action of microorganisms and the effects of biocides on carbonate rock of the Jeronimos Monastery (Lisbon)","volume":"49","author":"Ascaso","year":"2002","journal-title":"Int. Biodeterior. Biodegrad."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"11","DOI":"10.14568\/cp2012005","article-title":"Diversidade microbiol\u00f3gica do edif\u00edcio da Sacristia Incompleta do Convento de Cristo, em Tomar, e avalia\u00e7\u00e3o do seu controlo por biocidas","volume":"17","author":"Mateus","year":"2013","journal-title":"Conserv. Patrim\u00f3nio"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/0048-9697(95)04572-I","article-title":"Nature and decay effects of urban soiling on granitic building stones","volume":"167","author":"Schiavon","year":"1995","journal-title":"Sci. Total Environ."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1144\/GSL.SP.2002.205.01.15","article-title":"Biodeterioration of calcareous and granitic building stones in urban environments","volume":"205","author":"Schiavon","year":"2002","journal-title":"Geol. Soc. Lond. Spec. Publ."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"104734","DOI":"10.1016\/j.ibiod.2019.104734","article-title":"Deterioration and microbial colonization of cultural heritage stone buildings in polluted and unpolluted tropical and subtropical climates: A meta-analysis","volume":"143","author":"Neto","year":"2019","journal-title":"Int. Biodeterior. Biodegrad."},{"key":"ref_14","unstructured":"Battisti, V., and Gallo, V. (2019). A combined petrographic and geochemical metrological approach to assess the provenance of the building limestone used in the Batalha Monastery (Portugal). Metrology for Archaeology and Cultural Heritage, IMEKO."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"594","DOI":"10.3934\/microbiol.2018.4.594","article-title":"Natural limestone discolouration triggered by microbial activity\u2014A contribution","volume":"4","author":"Dias","year":"2018","journal-title":"AIMS Microbiol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1067","DOI":"10.1002\/mbo3.1030","article-title":"Assessment of microbiota present on a Portuguese historical stone convent using high-throughput sequencing approaches","volume":"9","author":"Rosado","year":"2020","journal-title":"MicrobiologyOpen"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1016\/j.culher.2019.07.025","article-title":"A change in composition, a change in colour: The case of limestone sculptures from the Portuguese National Museum of Ancient Art","volume":"42","author":"Dias","year":"2020","journal-title":"J. Cult. Herit."},{"key":"ref_18","first-page":"142","article-title":"An Updated Flora of Selmunett (St. Paul\u2019s Island) including Mosses and Lichens","volume":"4","author":"Mifsud","year":"2016","journal-title":"XJENZA"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0964-8305(02)00071-9","article-title":"Lichens and higher plants on stone: A review","volume":"51","author":"Lisci","year":"2003","journal-title":"Int. Biodeterior. Biodegrad."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1111\/j.1574-6968.1997.tb12620.x","article-title":"Solubilisation of some naturally occurring metal-bearing minerals, limescale and lead phosphate by Aspergillus niger","volume":"154","author":"Sayer","year":"1997","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1017\/S0953756297005510","article-title":"Solubilization of natural gypsum (CaSO4.2H2O) and the formation of calcium oxalate by Aspergillus niger and Serpula himantioides","volume":"102","author":"Gharieb","year":"1998","journal-title":"Mycol. Res."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/S0065-2164(08)00805-8","article-title":"Microbial deterioration of stone monuments\u2014an updated overview","volume":"66","author":"Scheerer","year":"2009","journal-title":"Adv. Appl. Microbiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"847","DOI":"10.5194\/acp-19-847-2019","article-title":"Phosphorus solubility in aerosol particles related to particle sources and atmospheric acidification in Asian continental outflow","volume":"19","author":"Shi","year":"2019","journal-title":"Atmos. Chem. Phys."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"fiw178","DOI":"10.1093\/femsec\/fiw178","article-title":"Intrinsic factors of Peltigera lichens influence the structure of the associated soil bacterial microbiota","volume":"92","author":"Leiva","year":"2016","journal-title":"FEMS Microbiol. Ecol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/j.1574-6968.2007.00918.x","article-title":"Bacterial endophytes: Recent developments and applications","volume":"278","author":"Ryan","year":"2008","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1309","DOI":"10.1128\/AEM.02257-10","article-title":"Bacterial communities associated with the lichen symbiosis","volume":"77","author":"Bates","year":"2011","journal-title":"Appl. Environ. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"428","DOI":"10.1134\/S0012496606050243","article-title":"The leading role of actinobacteria in aerobic cellulose degradation in Sphagnum peat bogs","volume":"410","author":"Pankratov","year":"2006","journal-title":"Dokl. Biol. Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"398","DOI":"10.3389\/fmicb.2020.00398","article-title":"The microbiomes of seven lichen genera reveal host specificity, a reduced core community and potential as source of antimicrobials","volume":"11","author":"Sierra","year":"2020","journal-title":"Front. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"434","DOI":"10.1111\/j.1758-2229.2010.00206.x","article-title":"Microbial metacommunities in the lichen\u2013rock habitat","volume":"3","author":"Bjelland","year":"2011","journal-title":"Environ. Microbiol. Rep."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"2711","DOI":"10.1007\/s11274-014-1695-z","article-title":"Biodiversity and physiological characteristics of Antarctic and Arctic lichens-associated bacteria","volume":"30","author":"Lee","year":"2014","journal-title":"World J. Microbiol. Biotechnol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.enggeo.2014.12.001","article-title":"Experimental studies on ignimbrite and the effect of lichens and capillarity on the deterioration of Seljuk Gravestones","volume":"185","author":"Oyan","year":"2015","journal-title":"Eng. Geol."}],"container-title":["Corrosion and Materials Degradation"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2624-5558\/2\/1\/2\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:10:37Z","timestamp":1760159437000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2624-5558\/2\/1\/2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,1,13]]},"references-count":31,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2021,3]]}},"alternative-id":["cmd2010002"],"URL":"https:\/\/doi.org\/10.3390\/cmd2010002","relation":{},"ISSN":["2624-5558"],"issn-type":[{"value":"2624-5558","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,1,13]]}}}