{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,8]],"date-time":"2026-05-08T15:32:41Z","timestamp":1778254361946,"version":"3.51.4"},"reference-count":96,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2020,8,10]],"date-time":"2020-08-10T00:00:00Z","timestamp":1597017600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2020,8,10]],"date-time":"2020-08-10T00:00:00Z","timestamp":1597017600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Marine biofouling remains an unsolved problem with a serious economic impact on several marine associated industries and constitutes a major vector for the spread of non-indigenous species (NIS). The implementation of biofouling monitoring programs allows for better fouling management and also for the early identification of NIS. However, few monitoring studies have used recent methods, such as metabarcoding, that can significantly enhance the detection of those species. Here, we employed monthly monitoring of biofouling growth on stainless steel plates in the Atlantic Port of Leix\u00f5es (Northern Portugal), over one year to test the effect of commercial anti-corrosion paint in the communities. Fouling organisms were identified by combining morpho-taxonomy identification with community DNA metabarcoding using multiple markers (16S rRNA, 18S rRNA, 23S rRNA, and COI genes). The dominant colonizers found at this location were hard foulers, namely barnacles and mussels, while other groups of organisms such as cnidarians, bryozoans, and ascidians were also abundant. Regarding the temporal dynamics of the fouling communities, there was a progressive increase in the colonization of cyanobacteria, green algae, and red algae during the sampled period with the replacement of less abundant groups. The tested anticorrosion paint demonstrated to have a significant prevention effect against the biofouling community resulting in a biomass reduction. Our study also reports, for the first time, 29 NIS in this port, substantiating the need for the implementation of recurring biofouling monitoring programs in ports and harbours.<\/jats:p>","DOI":"10.1038\/s41598-020-70307-4","type":"journal-article","created":{"date-parts":[[2020,8,10]],"date-time":"2020-08-10T10:03:08Z","timestamp":1597053788000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":45,"title":["Monitoring of biofouling communities in a Portuguese port using a combined morphological and metabarcoding approach"],"prefix":"10.1038","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3757-8143","authenticated-orcid":false,"given":"Joana","family":"Azevedo","sequence":"first","affiliation":[]},{"given":"Jorge T.","family":"Antunes","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6857-7581","authenticated-orcid":false,"given":"Andr\u00e9 M.","family":"Machado","sequence":"additional","affiliation":[]},{"given":"Vitor","family":"Vasconcelos","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5064-9164","authenticated-orcid":false,"given":"Pedro N.","family":"Le\u00e3o","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0262-0791","authenticated-orcid":false,"given":"Elsa","family":"Froufe","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2020,8,10]]},"reference":[{"key":"70307_CR1","doi-asserted-by":"crossref","first-page":"2879","DOI":"10.1111\/1462-2920.12186","volume":"15","author":"M Salta","year":"2013","unstructured":"Salta, M., Wharton, J. A., Blache, Y., Stokes, K. R. & Briand, J. F. Marine biofilms on artificial surfaces: Structure and dynamics. Environ. Microbiol.15, 2879\u20132893 (2013).","journal-title":"Environ. Microbiol."},{"key":"70307_CR2","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1111\/1758-2229.12694","volume":"11","author":"J Antunes","year":"2018","unstructured":"Antunes, J., Le\u00e3o, P. & Vasconcelos, V. Marine biofilms: diversity of communities and of chemical cues. Environ. Microbiol. Rep.11, 287\u2013305 (2018).","journal-title":"Environ. Microbiol. Rep."},{"key":"70307_CR3","doi-asserted-by":"crossref","DOI":"10.1201\/9780203503232","volume-title":"Marine Biofouling: Colonization Processes and Defenses","author":"AI Railkin","year":"2003","unstructured":"Railkin, A. I. Marine Biofouling: Colonization Processes and Defenses (CRC Press, Boca Raton, 2003)."},{"key":"70307_CR4","first-page":"164","volume":"23","author":"GW Swain","year":"2007","unstructured":"Swain, G. W., Kovach, B., Touzot, A. & Casse, F. K. C. Measuring the performance of today\u2019s antifouling coatings. J Sh. Prod23, 164\u2013170 (2007).","journal-title":"J Sh. Prod"},{"key":"70307_CR5","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1080\/08927014.2010.542809","volume":"27","author":"MP Schultz","year":"2011","unstructured":"Schultz, M. P., Bendick, J. A., Holm, E. R. & Hertel, W. M. Economic impact of biofouling on a naval surface ship. Biofouling27, 87\u201398 (2011).","journal-title":"Biofouling"},{"key":"70307_CR6","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1080\/08927014.2017.1320782","volume":"33","author":"L Holly Sweat","year":"2017","unstructured":"Holly Sweat, L., Swain, G. W. & K. Z. H. ,. Transported biofilms and their influence on subsequent macrofouling colonization. Biofouling33, 433\u2013449 (2017).","journal-title":"Biofouling"},{"key":"70307_CR7","doi-asserted-by":"crossref","first-page":"464","DOI":"10.1080\/08927014.2018.1464152","volume":"34","author":"MO Winfield","year":"2018","unstructured":"Winfield, M. O., Downer, A., Longyear, J., Dale, M. & Barker, G. L. A. Comparative study of biofilm formation on biocidal antifouling and fouling-release coatings using next-generation DNA sequencing. Biofouling34, 464\u2013477 (2018).","journal-title":"Biofouling"},{"key":"70307_CR8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41598-018-34447-y","volume":"8","author":"L Miralles","year":"2018","unstructured":"Miralles, L., Ardura, A., Clusa, L. & Garcia-Vazquez, E. DNA barcodes of Antipode marine invertebrates in Bay of Biscay and Gulf of Lion ports suggest new biofouling challenges. Sci. Rep.8, 1\u201311 (2018).","journal-title":"Sci. Rep."},{"key":"70307_CR9","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1016\/j.watres.2014.12.056","volume":"71","author":"J Jin","year":"2015","unstructured":"Jin, J., Wu, G. & Guan, Y. Effect of bacterial communities on the formation of cast iron corrosion tubercles in reclaimed water. Water Res.71, 207\u2013218 (2015).","journal-title":"Water Res."},{"key":"70307_CR10","doi-asserted-by":"crossref","unstructured":"Hou, B. R., et al. The cost of corrosion in China. H npj Mater. Degrad.1, (2017).","DOI":"10.1038\/s41529-017-0005-2"},{"key":"70307_CR11","doi-asserted-by":"crossref","first-page":"177","DOI":"10.3391\/mbi.2015.6.2.08","volume":"6","author":"T Riding","year":"2015","unstructured":"Riding, T. et al. The Marine Biosecurity Porthole \u2013 a web-based information system on non-indigenous marine species in New Zealand. Manag. Biol. Invasions6, 177\u2013184 (2015).","journal-title":"Manag. Biol. Invasions"},{"key":"70307_CR12","doi-asserted-by":"crossref","first-page":"20141517","DOI":"10.1098\/rspb.2014.1517","volume":"282","author":"FT Chan","year":"2015","unstructured":"Chan, F. T., Bradie, J., Briski, E., Bailey, S. A. & Simard, H. J. M. Assessing introduction risk using species\u2019 rank-abundance distributions. Proc. R. Soc. Biol. Sci.282, 20141517 (2015).","journal-title":"Proc. R. Soc. Biol. Sci."},{"key":"70307_CR13","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s00227-015-2782-x","volume":"163","author":"FT Chan","year":"2016","unstructured":"Chan, F. T., MacIsaac, H. J. & Bailey, S. A. Survival of ship biofouling assemblages during and after voyages to the Canadian Arctic. Mar. Biol.163, 1\u201314 (2016).","journal-title":"Mar. Biol."},{"issue":"2","key":"70307_CR14","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1111\/1755-0998.12971","volume":"19","author":"A Koziol","year":"2019","unstructured":"Koziol, A. et al. Environmental DNA metabarcoding studies are critically affected by substrate selection. Mol. Ecol. Resour.19(2), 66\u2013372 (2019).","journal-title":"Mol. Ecol. Resour."},{"key":"70307_CR15","doi-asserted-by":"crossref","first-page":"2452","DOI":"10.1002\/ece3.6071","volume":"10","author":"A Rey","year":"2020","unstructured":"Rey, A., Basurko, O. C. & Rodriguez-Ezpeleta, N. Considerations for metabarcoding-based port biological baseline surveys aimed at marine nonindigenous species monitoring and risk assessments. Ecol. Evol.10, 2452\u20132465 (2020).","journal-title":"Ecol. Evol."},{"key":"70307_CR16","doi-asserted-by":"crossref","first-page":"2045","DOI":"10.1111\/j.1365-294X.2012.05470.x","volume":"21","author":"P Taberlet","year":"2012","unstructured":"Taberlet, P., Coissac, E., Pompanon, F., Brochmann, C. & Willerslev, E. Towards next-generation biodiversity assessment using DNA metabarcoding. Mol. Ecol.21, 2045\u20132050 (2012).","journal-title":"Mol. Ecol."},{"key":"70307_CR17","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1080\/08927014.2015.1028923","volume":"31","author":"X Pochon","year":"2015","unstructured":"Pochon, X., Zaiko, A., Hopkins, G. A., Banks, J. C. & Wood, S. A. Early detection of eukaryotic communities from marine biofilm using high-throughput sequencing: an assessment of different sampling devices. Biofouling31, 241\u2013251 (2015).","journal-title":"Biofouling"},{"key":"70307_CR18","doi-asserted-by":"crossref","first-page":"16290","DOI":"10.1038\/s41598-018-34541-1","volume":"8","author":"UV Ammon","year":"2018","unstructured":"Ammon, U. V. et al. Combining morpho-taxonomy and metabarcoding enhances the detection of non-indigenous marine pests in biofouling communities. Sci Rep.8, 16290 (2018).","journal-title":"Sci Rep."},{"key":"70307_CR19","unstructured":"Slijkerman, D.M.E., et al. Monitoring Groningen Sea Ports. Wageningen, Wageningen Mar. Res. (2017)."},{"key":"70307_CR20","doi-asserted-by":"crossref","first-page":"e73935","DOI":"10.1371\/journal.pone.0073935","volume":"8","author":"X Pochon","year":"2013","unstructured":"Pochon, X., Bott, N. J., Smith, K. F. & Wood, S. Evaluating detection limits of next-generation sequencing for the surveillance and monitoring of international marine pests. PLoS ONE8, e73935 (2013).","journal-title":"PLoS ONE"},{"key":"70307_CR21","doi-asserted-by":"crossref","first-page":"1211","DOI":"10.1080\/08927014.2014.977267","volume":"30","author":"DH Leary","year":"2014","unstructured":"Leary, D. H. et al. Integrated metagenomic and metaproteomic analyses of marine biofilm communities. Biofouling30, 1211\u20131223 (2014).","journal-title":"Biofouling"},{"key":"70307_CR22","doi-asserted-by":"crossref","first-page":"657","DOI":"10.1080\/08927014.2018.1480757","volume":"34","author":"J-F Briand","year":"2018","unstructured":"Briand, J.-F. et al. Metabarcoding and metabolomics offer complementarity in deciphering marine eukaryotic biofouling community shifts. Biofouling34, 657\u2013672 (2018).","journal-title":"Biofouling"},{"key":"70307_CR23","doi-asserted-by":"crossref","first-page":"1295","DOI":"10.1016\/j.scitotenv.2018.05.002","volume":"637","author":"J Pawlowski","year":"2018","unstructured":"Pawlowski, J. et al. The future of biotic indices in the ecogenomic era: Integrating (e)DNA metabarcoding in biological assessment of aquatic ecosystems. Sci. Total Environ.637, 1295\u20131310 (2018).","journal-title":"Sci. Total Environ."},{"key":"70307_CR24","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1016\/j.ecolind.2018.07.044","volume":"95","author":"E Aylagas","year":"2018","unstructured":"Aylagas, E., Borja, \u00c1, Muxika, I. & Rodr\u00edguez-Ezpeleta, N. Adapting metabarcoding-based benthic biomonitoring into routine marine ecological status assessment networks. Ecol. Indic.95, 194\u2013202 (2018).","journal-title":"Ecol. Indic."},{"key":"70307_CR25","doi-asserted-by":"crossref","first-page":"11559","DOI":"10.1038\/s41598-019-47899-7","volume":"9","author":"LE Holman","year":"2019","unstructured":"Holman, L. E. et al. Detection of introduced and resident marine species using environmental DNA metabarcoding of sediment and water. Sci. Rep.9, 11559 (2019).","journal-title":"Sci. Rep."},{"key":"70307_CR26","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1111\/1755-0998.12982","volume":"19","author":"GJ Jeunen","year":"2019","unstructured":"Jeunen, G. J. et al. Environmental DNA (eDNA) metabarcoding reveals strong discrimination among diverse marine habitats connected by water movement. Mol. Ecol. Resour.19, 426\u2013438 (2019).","journal-title":"Mol. Ecol. Resour."},{"key":"70307_CR27","doi-asserted-by":"crossref","first-page":"e0183347","DOI":"10.1371\/journal.pone.0183347","volume":"12","author":"YJ Borrell","year":"2017","unstructured":"Borrell, Y. J., Miralles, L., Do Huu, H., Mohammed-Geba, K. & Garcia-Vazquez, E. DNA in a bottle: rapid metabarcoding survey for early alerts of invasive species in ports. PLoS ONE12, e0183347 (2017).","journal-title":"PLoS ONE"},{"key":"70307_CR28","doi-asserted-by":"crossref","first-page":"8843","DOI":"10.1038\/s41598-018-27048-2","volume":"8","author":"EK Grey","year":"2018","unstructured":"Grey, E. K. et al. Effects of sampling effort on biodiversity patterns estimated from environmental DNA metabarcoding surveys. Sci. Rep.8, 8843 (2018).","journal-title":"Sci. Rep."},{"key":"70307_CR29","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1080\/08927014.2016.1186165","volume":"32","author":"A Zaiko","year":"2016","unstructured":"Zaiko, A. et al. Metabarcoding improves detection of eukaryotes from early biofouling communities: implications for pest monitoring and pathway management. Biofouling32, 671\u2013684 (2016).","journal-title":"Biofouling"},{"key":"70307_CR30","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/j.marenvres.2017.12.003","volume":"133","author":"U von Ammon","year":"2018","unstructured":"von Ammon, U. et al. The impact of artificial surfaces on marine bacterial and eukaryotic biofouling assemblages: a high-throughput sequencing analysis. Mar. Environ. Res.133, 57\u201366 (2018).","journal-title":"Mar. Environ. Res."},{"key":"70307_CR31","unstructured":"MSFD, Directive 2008\/56\/EC."},{"key":"70307_CR32","doi-asserted-by":"crossref","first-page":"54","DOI":"10.1016\/j.marpol.2017.08.014","volume":"85","author":"JA Darling","year":"2017","unstructured":"Darling, J. A. et al. Recommendations for developing and applying molecular genetic tools to assess and manage biological invasions in marine ecosystems. Mar Policy85, 54\u201364 (2017).","journal-title":"Mar Policy"},{"key":"70307_CR33","unstructured":"IMO, 2004. International Convention on the Control and Management of Ships\u2019 Ballast Water and Sediments. International Maritime Organisation, London (www.imo.org)."},{"key":"70307_CR34","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.marpolbul.2010.09.014","volume":"62","author":"MJ Bishop","year":"2011","unstructured":"Bishop, M. J. & Hutchings, P. A. How useful are port surveys focused on target pest identification for exotic species management?. Mar. Pollut. Bull.62, 36\u201342 (2011).","journal-title":"Mar. Pollut. Bull."},{"key":"70307_CR35","unstructured":"Awad, A., Haag, F., Anil, A.C., Abdulla, A. GEF-UNDP-IMO GloBallast Partnerships Programme, IOI, CSIR-NIO and IUCN. Guidance on Port Biological Baseline Surveys. GEF-UNDP-IMO GloBallast Partnerships, London, UK. GloBallast Monograph No. 22. (2014)."},{"key":"70307_CR36","doi-asserted-by":"crossref","first-page":"199","DOI":"10.1016\/j.ecss.2015.06.019","volume":"167","author":"P Chainho","year":"2015","unstructured":"Chainho, P. et al. Non-indigenous species in Portuguese coastal areas, coastal lagoons, estuaries and islands. Estuar. Coast. Shelf Sci.167, 199\u2013211 (2015).","journal-title":"Estuar. Coast. Shelf Sci."},{"key":"70307_CR37","doi-asserted-by":"crossref","first-page":"62","DOI":"10.1016\/j.jenvman.2019.01.039","volume":"235","author":"RAO Nunes","year":"2019","unstructured":"Nunes, R. A. O., Alvim-Ferraz, M. C. M., Martins, F. G. & Sousa, S. I. V. Environmental and social valuation of shipping emissions on four ports of Portugal. J. Environ. Manag.235, 62\u201369 (2019).","journal-title":"J. Environ. Manag."},{"key":"70307_CR38","first-page":"89","volume":"21","author":"I Menchaca","year":"2014","unstructured":"Menchaca, I. et al. Guide for the evaluation of biofouling formation in the marine environment. Rev. Investig. Mar. AZTI-Tecnalia21, 89\u201399 (2014).","journal-title":"Rev. Investig. Mar. AZTI-Tecnalia"},{"key":"70307_CR39","unstructured":"ASTM. D3623 \u2212 78a (Reapproved 2012) Standard Test Method for Testing Antifouling Panels in Shallow Submergence. ASTM International (2012)."},{"key":"70307_CR40","doi-asserted-by":"crossref","first-page":"261","DOI":"10.3391\/ai.2013.8.3.02","volume":"8","author":"J Canning-Clode","year":"2014","unstructured":"Canning-Clode, J. et al. Marine invasions on a subtropical island: fouling studies and new records in a recent marina on Madeira Island (Eastern Atlantic Ocean). Aquat. Invasions8, 261\u2013270 (2014).","journal-title":"Aquat. Invasions"},{"key":"70307_CR41","doi-asserted-by":"crossref","first-page":"153","DOI":"10.1007\/s00248-006-9154-5","volume":"53","author":"PR Jones","year":"2007","unstructured":"Jones, P. R., Cottrell, M. T., Kirchman, D. L. & Dexter, S. C. Bacterial community structure of biofilms on artificial surfaces in an estuary. Microb. Ecol.53, 153\u2013162 (2007).","journal-title":"Microb. Ecol."},{"key":"70307_CR42","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1007\/s12275-008-0032-3","volume":"46","author":"JW Lee","year":"2008","unstructured":"Lee, J. W., Nam, J. H., Kim, Y. H., Lee, K. H. & Lee, D. H. Bacterial communities in the initial stage of marine biofilm formation on artificial surfaces. J. Microbiol.46, 174\u2013182 (2008).","journal-title":"J. Microbiol."},{"key":"70307_CR43","unstructured":"Kraberg, A. et al. Coastal Phytoplankton: Photo Guide for Northern European Seas. Edition: 1 Publisher: Pfeil Verlag Editor: Karen H. Wiltshire, Maarten Boersma. (Pfeil Verlag, 2010)."},{"key":"70307_CR44","unstructured":"Hoppenrath M. et al. Marine Phytoplankton Selected microphytoplankton species from the North Sea around Helgoland and Sylt. 264 pages, 87 figures. (2009)."},{"key":"70307_CR45","unstructured":"Mees, J. et al. World Register of Marine Species. Available from http:\/\/www.marinespecies.org (2015)."},{"key":"#cr-split#-70307_CR46.1","unstructured":"Guiry, M. D. & Guiry, G. M. AlgaeBase. World-wide electronic publication, National University of Ireland, Galway. https:\/\/www.algaebase.org; searched on 21 March 2017. AlgaeBase. World-wide electronic publication, National University of Ireland, Galway."},{"key":"#cr-split#-70307_CR46.2","unstructured":"https:\/\/www.algaebase.org; searched on 21 March 2017. (2017)."},{"key":"70307_CR47","doi-asserted-by":"crossref","first-page":"671","DOI":"10.1038\/nmeth.2089","volume":"9","author":"CA Schneider","year":"2012","unstructured":"Schneider, C. A., Rasband, W. S. & Eliceiri, K. W. NIH Image to ImageJ: 25 years of image analysis. Nat. Methods9, 671 (2012).","journal-title":"Nat. Methods"},{"key":"70307_CR48","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","volume":"9","author":"J Shindelin","year":"2012","unstructured":"Shindelin, J. et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods9, 676\u2013682 (2012).","journal-title":"Nat. Methods"},{"key":"70307_CR49","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1093\/nar\/gks1039","volume":"41","author":"AJ Sabat","year":"2013","unstructured":"Sabat, A. J. et al. Evaluation of general 16S ribosomal RNA gene PCR primers for classical and next-generation sequencing-based diversity studies. Nucleic. Acids Res.41, 1 (2013).","journal-title":"Nucleic. Acids Res."},{"key":"70307_CR50","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1111\/j.1365-294X.2009.04480.x","volume":"19","author":"T Stoeck","year":"2010","unstructured":"Stoeck, T., Bass, D., Nebel, M., Christen, R. & Meredith, D. Multiple marker parallel tag environmental DNA sequencing reveals a highly complex eukaryotic community in marine anoxic water. Mol. Ecol.19, 21\u201331 (2010).","journal-title":"Mol. Ecol."},{"key":"70307_CR51","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1186\/1742-9994-10-34","volume":"10","author":"M Leray","year":"2013","unstructured":"Leray, M. et al. A new versatile primer set targeting a short fragment of the mitochondrial COI region for metabarcoding metazoan diversity: application for characterizing coral reef fish gut contents. Front. Zool.10, 34 (2013).","journal-title":"Front. Zool."},{"key":"70307_CR52","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1046\/j.1095-8312.2003.00197.x","volume":"79","author":"CP Meyer","year":"2003","unstructured":"Meyer, C. P. Molecular systematics of cowries (Gastropoda: Cypraeidae) and diversification patterns in the tropics. Biol. J. Linn. Soc.79, 401\u2013459 (2003).","journal-title":"Biol. J. Linn. Soc."},{"key":"70307_CR53","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1111\/j.1529-8817.2007.00341.x","volume":"43","author":"AR Sherwood","year":"2007","unstructured":"Sherwood, A. R. & Presting, G. G. Universal primers amplify a 23S rDNA plastid marker in eukaryotic algae and cyanobacteria. J. Phycol.43, 605\u2013608 (2007).","journal-title":"J. Phycol."},{"issue":"5","key":"70307_CR54","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1038\/nmeth.f.303","volume":"7","author":"JG Caporaso","year":"2010","unstructured":"Caporaso, J. G. et al. QIIME allows analysis of high-throughput community sequencing data. Nat. Methods7(5), 335\u2013336 (2010).","journal-title":"Nat. Methods"},{"key":"70307_CR55","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1038\/nmeth.2276","volume":"10","author":"NA Bokulich","year":"2013","unstructured":"Bokulich, N. A. et al. Quality-filtering vastly improves diversity estimates from Illumina amplicon sequencing. Nat. Methods10, 57\u201359 (2013).","journal-title":"Nat. Methods"},{"key":"70307_CR56","doi-asserted-by":"crossref","first-page":"2194","DOI":"10.1093\/bioinformatics\/btr381","volume":"16","author":"RC Edgar","year":"2011","unstructured":"Edgar, R. C., Haas, B. J., Clemente, J. C., Quince, C. & Knight, R. UCHIME improves sensitivity and speed of chimera detection. Bioinformatics16, 2194\u20132200 (2011).","journal-title":"Bioinformatics"},{"key":"70307_CR57","doi-asserted-by":"crossref","first-page":"996","DOI":"10.1038\/nmeth.2604","volume":"10","author":"RC Edgar","year":"2013","unstructured":"Edgar, R. C. UPARSE: Highly accurate OTU sequences from microbial amplicon reads. Nat. Methods10, 996\u2013998 (2013).","journal-title":"Nat. Methods"},{"key":"70307_CR58","doi-asserted-by":"crossref","first-page":"5069","DOI":"10.1128\/AEM.03006-05","volume":"72","author":"TZ DeSantis","year":"2006","unstructured":"DeSantis, T. Z. et al. Greengenes, a chimera-checked 16S rRNA gene database and workbench compatible with ARB. Appl. Environ. Microbiol.72, 5069\u20135072 (2006).","journal-title":"Appl. Environ. Microbiol."},{"key":"70307_CR59","doi-asserted-by":"crossref","first-page":"D590","DOI":"10.1093\/nar\/gks1219","volume":"41","author":"C Quast","year":"2013","unstructured":"Quast, C. et al. The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res.41, D590\u2013D596 (2013).","journal-title":"Nucleic Acids Res."},{"key":"70307_CR60","doi-asserted-by":"crossref","first-page":"5261","DOI":"10.1128\/AEM.00062-07","volume":"73","author":"Q Wang","year":"2007","unstructured":"Wang, Q., Garrity, G. M., Tiedje, J. M. & Cole, J. R. Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl. Environ. Microbiol.73, 5261\u20135267 (2007).","journal-title":"Appl. Environ. Microbiol."},{"key":"70307_CR61","doi-asserted-by":"crossref","unstructured":"Ratnasingham, S. & Herbert, P. D. N. bold: The Barcode of Life Data System (https:\/\/www.barcodinglife.org). Mol. Ecol. Notes7, 355\u2013364 (2007).","DOI":"10.1111\/j.1471-8286.2007.01678.x"},{"key":"70307_CR62","doi-asserted-by":"crossref","first-page":"426","DOI":"10.1093\/bioinformatics\/btg430","volume":"20","author":"M Clamp","year":"2004","unstructured":"Clamp, M., Cuff, J., Searle, S. M. & Barton, G. J. The Jalview Java alignment editor. Bioinformatics20, 426\u2013427 (2004).","journal-title":"Bioinformatics"},{"key":"70307_CR63","doi-asserted-by":"crossref","first-page":"W7","DOI":"10.1093\/nar\/gkq291","volume":"38","author":"F Abascal","year":"2010","unstructured":"Abascal, F., Zardoya, R. & Telford, M. J. TranslatorX: multiple alignment of nucleotide sequences guided by amino acid translations. Nucleic Acids Res.38, W7\u2013W13 (2010).","journal-title":"Nucleic Acids Res."},{"issue":"4","key":"70307_CR64","doi-asserted-by":"crossref","first-page":"e61217","DOI":"10.1371\/journal.pone.0061217","volume":"8","author":"PJ McMurdie","year":"2013","unstructured":"McMurdie, P. J. & Holmes, S. Phyloseq: An R package for reproducible interactive analysis and graphics of microbiome census data. PLoS ONE8(4), e61217 (2013).","journal-title":"PLoS ONE"},{"key":"70307_CR65","unstructured":"Oksanen, J. et al. vegan: Community Ecology Package. R package version 2.5-2. Cran R (2019)."},{"key":"70307_CR66","unstructured":"RStudio Team. RStudio: Integrated Development for R [Computer software]. RStudio, Inc. (2015)."},{"key":"70307_CR67","first-page":"32","volume":"26","author":"MJ Anderson","year":"2001","unstructured":"Anderson, M. J. New method for non-parametric multi- variate analysis of variance. Austral. Ecol.26, 32\u201346 (2001).","journal-title":"Austral. Ecol."},{"key":"70307_CR68","unstructured":"Invasive Species Specialist Group ISSG. The Global Invasive Species Database. Version 2015.1 (2015)."},{"key":"70307_CR69","unstructured":"Palomares, M. L. D. & Pauly, D. SeaLifeBase, version (08\/2019). World Wide Web electronic publication (2019)."},{"key":"70307_CR70","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1017\/S002531540501132Xh","volume":"85","author":"MH Davis","year":"2005","unstructured":"Davis, M. H. & Davis, M. E. Styela clava (Tunicata: Ascidiacea): a new addition to the fauna of the Portuguese coast. J. Mar. Biol. Assoc. UK85, 403\u2013404 (2005).","journal-title":"J. Mar. Biol. Assoc. UK"},{"key":"70307_CR71","doi-asserted-by":"crossref","first-page":"378","DOI":"10.3391\/ai.2007.2.4.6","volume":"2","author":"MH Davis","year":"2007","unstructured":"Davis, M. H., L\u00fctzen, J. & Davis, M. E. The spread of Styela clava Herdman, 1882 (Tunicata, Ascidiacea) in European waters. Aquat. Invasions2, 378\u2013390 (2007).","journal-title":"Aquat. Invasions"},{"key":"70307_CR72","doi-asserted-by":"crossref","first-page":"929","DOI":"10.1111\/mec.13428","volume":"25","author":"A Valentini","year":"2015","unstructured":"Valentini, A. et al. Next-generation monitoring of aquatic biodiversity using environmental DNA metabarcoding. Mol. Ecol.25, 929\u2013942 (2015).","journal-title":"Mol. Ecol."},{"key":"70307_CR73","doi-asserted-by":"crossref","first-page":"6918","DOI":"10.1002\/ece3.3192","volume":"7","author":"V Elbrecht","year":"2017","unstructured":"Elbrecht, V., Peinert, B. & Leese, F. Sorting things out: assessing effects of unequal specimen biomass on DNA metabarcoding. Ecol. Evol.7, 6918\u20136926 (2017).","journal-title":"Ecol. Evol."},{"key":"70307_CR74","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1007\/s00227-016-2911-1","volume":"163","author":"W Xiong","year":"2016","unstructured":"Xiong, W., Li, H. & Zhan, A. Early detection of invasive species in marine ecosystems using high-throughput sequencing: technical challenges and possible solutions. Mar. Biol.163, 1\u201312 (2016).","journal-title":"Mar. Biol."},{"key":"70307_CR75","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1111\/1755-0998.12338","volume":"15","author":"GF Ficetola","year":"2015","unstructured":"Ficetola, G. F. et al. Replication levels, false presences and the estimation of the presence\/absence from eDNA metabarcoding data. Mol. Ecol. Resour.15, 543\u2013556 (2015).","journal-title":"Mol. Ecol. Resour."},{"key":"70307_CR76","doi-asserted-by":"crossref","first-page":"2252","DOI":"10.1002\/ece3.1497","volume":"5","author":"JM Flynn","year":"2015","unstructured":"Flynn, J. M., Brown, E. A., Chain, F. J. J., Macisaac, H. J. & Cristescu, M. E. Toward accurate molecular identification of species in complex environmental samples: Testing the performance of sequence filtering and clustering methods. Ecol. Evol.5, 2252\u20132266 (2015).","journal-title":"Ecol. Evol."},{"key":"70307_CR77","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1016\/j.culher.2008.07.010","volume":"10","author":"CJ McNamara","year":"2009","unstructured":"McNamara, C. J., Bearce Lee, K., Russell, M. A., Murphy, L. E. & Mitchell, R. Analysis of bacterial community composition in concretions formed on the USS Arizona, Pearl Harbor. HI. J. Cult. Herit10, 232\u2013236 (2009).","journal-title":"HI. J. Cult. Herit"},{"key":"70307_CR78","doi-asserted-by":"crossref","first-page":"4821","DOI":"10.1128\/AEM.00948-14","volume":"80","author":"M Camps","year":"2014","unstructured":"Camps, M. et al. Antifouling coatings influence both abundance and community structure of colonizing biofilms: A case study in the Northwestern Mediterranean Sea. Appl. Environ. Microbiol.80, 4821\u20134831 (2014).","journal-title":"Appl. Environ. Microbiol."},{"key":"70307_CR79","doi-asserted-by":"crossref","first-page":"1155","DOI":"10.1080\/08927014.2014.972951","volume":"30","author":"T Muthukrishnan","year":"2014","unstructured":"Muthukrishnan, T., Abed, R. M. M., Dobretsov, S., Kidd, B. & Finnie, A. Long-term microfouling on commercial biocidal fouling control coatings. Biofouling30, 1155\u20131164 (2014).","journal-title":"Biofouling"},{"key":"70307_CR80","doi-asserted-by":"crossref","first-page":"585","DOI":"10.1007\/s00248-017-0966-2","volume":"74","author":"JF Briand","year":"2017","unstructured":"Briand, J. F. et al. Spatio-temporal variations of marine biofilm communities colonizing artificial substrata including antifouling coatings in contrasted french coastal environments. Microb. Ecol.74, 585\u2013598 (2017).","journal-title":"Microb. Ecol."},{"key":"70307_CR81","first-page":"6","volume":"94","author":"T Pollet","year":"2019","unstructured":"Pollet, T. et al. Prokaryotic community successions and interactions in marine biofilms: the key role of Flavobacteriia. FEMS Microbiol. Ecol.94, 6 (2019).","journal-title":"FEMS Microbiol. Ecol."},{"key":"70307_CR82","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1128\/MMBR.00037-15","volume":"80","author":"H Dang","year":"2016","unstructured":"Dang, H. & Lovell, C. R. Microbial surface colonization and biofilm development in marine environments. Microbiol. Mol. Biol. Rev.80, 91\u2013138 (2016).","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"70307_CR83","first-page":"91","volume":"39","author":"DL Kirchman","year":"2002","unstructured":"Kirchman, D. L. The ecology of Cytophaga-Flavobacteria in aquatic environments. FEMS Microbiol. Ecol.39, 91\u2013100 (2002).","journal-title":"FEMS Microbiol. Ecol."},{"key":"70307_CR84","doi-asserted-by":"crossref","first-page":"467","DOI":"10.1128\/AEM.66.2.467-475.2000","volume":"66","author":"H Dang","year":"2000","unstructured":"Dang, H. & Lovell, C. R. Bacterial primary colonization and early succession on surfaces in marine waters as determined by amplified rRNA gene restriction analysis and sequence analysis of 16S rRNA genes. Appl. Environ. Microbiol.66, 467\u2013475 (2000).","journal-title":"Appl. Environ. Microbiol."},{"key":"70307_CR85","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1016\/j.marpolbul.2014.12.003","volume":"91","author":"S Saloni","year":"2015","unstructured":"Saloni, S. & Crowe, T. P. Impacts of multiple stressors during the establishment of fouling assemblages. Mar. Pollut. Bull.91, 211\u2013221 (2015).","journal-title":"Mar. Pollut. Bull."},{"key":"70307_CR86","doi-asserted-by":"crossref","first-page":"103","DOI":"10.3354\/meps226103","volume":"226","author":"EL Johnston","year":"2002","unstructured":"Johnston, E. L., Keough, M. J. & Qian, P.-Y. Maintenance of species dominance through pulse disturbances to a sessile marine invertebrate assemblage in Port Shelter, Hong Kong. Mar. Ecol. Prog. Ser.226, 103\u2013114 (2002).","journal-title":"Mar. Ecol. Prog. Ser."},{"key":"70307_CR87","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.jembe.2011.04.001","volume":"403","author":"C Antoniadou","year":"2011","unstructured":"Antoniadou, C., Voultsiadou, E. & Chintiroglou, C. Seasonal patterns of colonization and early succession on sublittoral rocky cliffs. J. Exp. Mar. Biol. Ecol.403, 21\u201330 (2011).","journal-title":"J. Exp. Mar. Biol. Ecol."},{"key":"70307_CR88","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1007\/s00338-006-0191-9","volume":"26","author":"SN Field","year":"2007","unstructured":"Field, S. N., Glassom, D. & Bythell, J. Effects of artificial settlement plate materials and methods of deployment on the sessile epibenthic community development in a tropical environment. Coral Reefs26, 279\u2013289 (2007).","journal-title":"Coral Reefs"},{"key":"70307_CR89","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.jembe.2010.08.019","volume":"395","author":"M Cifuentes","year":"2010","unstructured":"Cifuentes, M., Krueger, I., Dumont, C. P., Lenz, M. & Thiel, M. Does primary colonization or community structure determine the succession of fouling communities?. J. Exp. Mar. Biol. Ecol.395, 10\u201320 (2010).","journal-title":"J. Exp. Mar. Biol. Ecol."},{"key":"70307_CR90","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1046\/j.1439-0485.2001.01729.x","volume":"22","author":"I Svane","year":"2001","unstructured":"Svane, I. & Petersen, J. K. On the problems of epibioses, fouling and artificial reefs, a review. Mar. Ecol.22, 169\u2013188 (2001).","journal-title":"Mar. Ecol."},{"key":"70307_CR91","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1016\/j.jembe.2005.12.029","volume":"330","author":"AJ Underwood","year":"2006","unstructured":"Underwood, A. J. & Chapman, M. G. Early development of subtidal macrofaunal assemblages: Relationships to period and timing of colonization. J. Exp. Mar. Biol. Ecol.330, 221\u2013233 (2006).","journal-title":"J. Exp. Mar. Biol. Ecol."},{"key":"70307_CR92","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1111\/j.1439-0485.2010.00398.x","volume":"32","author":"AS Pacheco","year":"2011","unstructured":"Pacheco, A. S., Laudien, J., Thiel, M., Oliva, M. & Heilmayer, O. Succession and seasonal onset of colonization in subtidal hard-bottom communities off northern Chile. Mar. Ecol.32, 75\u201387 (2011).","journal-title":"Mar. Ecol."},{"key":"70307_CR93","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3354\/meps316001","volume":"316","author":"DA Bowden","year":"2006","unstructured":"Bowden, D. A., Clarke, A., Peck, L. S. & Barnes, D. K. A. Antarctic sessile marine benthos: Colonisation and growth on artificial substrata over three years. Mar. Ecol. Prog. Ser.316, 1\u201316 (2006).","journal-title":"Mar. Ecol. Prog. Ser."},{"key":"70307_CR94","first-page":"46","volume":"40","author":"C Hewitt","year":"2001","unstructured":"Hewitt, C. & Martin, R. Revised protocols for baseline port surveys for introduced marine species: survey design, sampling protocols and specimen handling. CRIMP Tech. Rep.40, 46 (2001).","journal-title":"CRIMP Tech. Rep."},{"key":"70307_CR95","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1007\/s00227-016-2920-0","volume":"163","author":"D Abad","year":"2016","unstructured":"Abad, D. et al. Is metabarcoding suitable for estuarine plankton monitoring? A comparative study with microscopy. Mar. Biol.163, 149 (2016).","journal-title":"Mar. Biol."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-70307-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-70307-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-70307-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,5]],"date-time":"2023-10-05T14:21:17Z","timestamp":1696515677000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-70307-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,8,10]]},"references-count":96,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2020,12]]}},"alternative-id":["70307"],"URL":"https:\/\/doi.org\/10.1038\/s41598-020-70307-4","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,8,10]]},"assertion":[{"value":"14 June 2019","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"15 June 2020","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 August 2020","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"13461"}}