{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:02:47Z","timestamp":1785402167994,"version":"3.56.0"},"reference-count":59,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T00:00:00Z","timestamp":1777507200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Institutional Grant number 565 of Military Institute of Medicine\u2013National Research Institute"},{"id":[{"id":"https:\/\/ror.org\/05bts0a67","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Odontogenic sinusitis (ODS) is a common cause of unilateral maxillary sinusitis arising from periapical lesions (PALs) or other dental sources. The infection is typically polymicrobial and dominated by anaerobic bacteria, which are often under detected by routine culture. Molecular approaches such as quantitative polymerase chain reaction (QPCR) and next-generation sequencing (NGS) may provide improved characterization of the microbial burden and community structure. This study aimed to compare culture-based methods, targeted quantitative PCR, and 16S rRNA sequencing in paired samples to characterize microbial composition of ODS and evaluate diagnostic performance. Paired sinus mucosal biopsy (SIN) and periapical lesion (PAL) samples were collected from 28 patients with clinically confirmed ODS. Bacterial detection was performed using conventional culture and targeted QPCR assays for ten clinically relevant taxa. In three randomly selected patients, paired samples were additionally analyzed by 16S rRNA gene amplicon sequencing. Microbial load, taxa richness, and similarity between the two anatomically connected sites were assessed using Wilcoxon signed-rank, McNemar, Jaccard distance, and Bray\u2013Curtis dissimilarity analyses. Results: Culture showed low sensitivity, identifying a limited number of pathogens, primarily Staphylococcus aureus, Streptococcus anginosus, and Fusobacterium nucleatum, in a minority of samples. In contrast, QPCR demonstrated substantially higher detection rates, particularly in PAL samples. Porphyromonas gingivalis (96.8%), Fusobacterium spp. (100.0%), and the S. anginosus group (90.3%) were highly prevalent in PAL specimens, with overlapping but lower detection in SIN samples. PAL samples exhibited significantly higher bacterial loads and taxa richness than paired SIN samples (Wilcoxon p = 0.0004). 16S rRNA sequencing confirmed polymicrobial communities at both sites and identified additional taxa not included in the QPCR panel. Similarity analyses revealed pronounced interindividual variability, ranging from near-identical to highly divergent paired microbiota. Periapical lesions act as reservoirs of predominantly anaerobic bacteria that may seed the maxillary sinus in ODS. Although microbial overlap exists, sinus communities display lower burden and site-specific compositional shifts. Culture-based diagnostics underestimate ODS microbial complexity, whereas combined molecular approaches provide a more comprehensive and clinically informative assessment.<\/jats:p>","DOI":"10.3390\/ijms27094010","type":"journal-article","created":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T10:47:28Z","timestamp":1777546048000},"page":"4010","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Quantitative PCR-Based Analysis of Bacterial Profiles in Periapical Lesions and Maxillary Sinus in Odontogenic Sinusitis"],"prefix":"10.3390","volume":"27","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3683-9508","authenticated-orcid":false,"given":"Marta Aleksandra","family":"Kwiatkowska","sequence":"first","affiliation":[{"name":"Department of Otolaryngology and Oncological Laryngology with Division of Cranio-Maxillo-Facial Surgery, Military Institute of Medicine\u2013National Research Institute, 04-141 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7847-049X","authenticated-orcid":false,"given":"Alicja","family":"Tr\u0119bi\u0144ska-Stryjewska","sequence":"additional","affiliation":[{"name":"Biomedical Engineering Centre, Institute of Optoelectronics, Military University of Technology, 00-908 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katarzyna","family":"Andrejuk","sequence":"additional","affiliation":[{"name":"Biomedical Engineering Centre, Institute of Optoelectronics, Military University of Technology, 00-908 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dariusz","family":"Jurkiewicz","sequence":"additional","affiliation":[{"name":"Department of Otolaryngology and Oncological Laryngology with Division of Cranio-Maxillo-Facial Surgery, Military Institute of Medicine\u2013National Research Institute, 04-141 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0318-1698","authenticated-orcid":false,"given":"El\u017cbieta Anna","family":"Trafny","sequence":"additional","affiliation":[{"name":"Biomedical Engineering Centre, Institute of Optoelectronics, Military University of Technology, 00-908 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6394-8451","authenticated-orcid":false,"given":"Aneta","family":"Guzek","sequence":"additional","affiliation":[{"name":"Department of Laboratory Diagnostics, Section of Microbiology, Military Institute of Medicine\u2013National Research Institute, 04-141 Warsaw, Poland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1002\/wjo2.9","article-title":"Odontogenic sinusitis: A state-of-the-art review","volume":"8","author":"Craig","year":"2022","journal-title":"World J. Otorhinolaryngol. Head Neck Surg."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"349","DOI":"10.1016\/j.otohns.2005.10.059","article-title":"Sinusitis of odontogenic origin","volume":"135","author":"Brook","year":"2006","journal-title":"Otolaryngol. Head Neck Surg."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1002\/lio2.147","article-title":"Odontogenic sinusitis: A review of the current literature","volume":"3","author":"Little","year":"2018","journal-title":"Laryngoscope Investig. Otolaryngol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1353","DOI":"10.1099\/jmm.0.031476-0","article-title":"Bacteriological findings and antimicrobial resistance in odontogenic and non-odontogenic chronic maxillary sinusitis","volume":"60","author":"Puglisi","year":"2011","journal-title":"J. Med. Microbiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1007","DOI":"10.1016\/j.otc.2024.06.006","article-title":"Sinus Pathophysiology of Odontogenic Sinusitis","volume":"57","author":"Craig","year":"2024","journal-title":"Otolaryngol. Clin. N. Am."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1241","DOI":"10.30574\/wjarr.2023.18.1.0757","article-title":"Odontogenic sinusitis, a differential diagnosis: Literature review","volume":"18","author":"Micaela","year":"2023","journal-title":"World J. Adv. Res. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"27","DOI":"10.1097\/MOO.0000000000000430","article-title":"Odontogenic sinusitis: Developments in diagnosis, microbiology, and treatment","volume":"26","author":"Workman","year":"2018","journal-title":"Curr. Opin. Otolaryngol. Head Neck Surg."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1007\/s00784-024-06097-0","article-title":"Microbial spectrum and resistance of odontogenic abscesses\u2014Microbiological analysis using next generation sequencing","volume":"29","author":"Thol","year":"2025","journal-title":"Clin. Oral Investig."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1016\/j.wjorl.2018.11.001","article-title":"Determining the utility of standard hospital microbiology testing: Comparing standard microbiology cultures with DNA sequence analysis in patients with chronic sinusitis","volume":"5","author":"Rapoport","year":"2019","journal-title":"World J. Otorhinolaryngol. Head Neck Surg."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1016\/j.otc.2024.06.003","article-title":"Complicated Odontogenic Sinusitis: Extrasinus Infectious Spread","volume":"57","author":"Yu","year":"2024","journal-title":"Otolaryngol. Clin. N. Am."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e21","DOI":"10.1016\/j.medin.2017.07.003","article-title":"Subdural empyema as a complication of odontogenic maxillary sinusitis","volume":"42","year":"2018","journal-title":"Med. Intensiv."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.otot.2017.08.005","article-title":"Orbital complications of acute sinusitis","volume":"28","author":"Campbell","year":"2017","journal-title":"Oper. Tech. Otolaryngol. Head Neck Surg."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1016\/j.jcms.2017.01.023","article-title":"Odontogenic sinusitis maxillaris: A retrospective study of 121 cases with surgical intervention","volume":"45","author":"Zirk","year":"2017","journal-title":"J. Cranio-Maxillofac. Surg."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"111267","DOI":"10.1016\/j.humimm.2025.111267","article-title":"Comprehensive profiling of the heterogeneity of molecular endotypic traits in chronic rhinosinusitis","volume":"86","author":"Takahashi","year":"2025","journal-title":"Hum. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"B\u00f6ttger, S., Zechel-Gran, S., Schmermund, D., Streckbein, P., Wilbrand, J.F., Knitschke, M., Pons-K\u00fchnemann, J., Hain, T., Weigel, M., and Imirzalioglu, C. (2021). Clinical relevance of the microbiome in odontogenic abscesses. Biology, 10.","DOI":"10.3390\/biology10090916"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1150","DOI":"10.1128\/JCM.02082-09","article-title":"Development of Real-Time PCR Assays for Detection of the Streptococcus milleri Group from Cystic Fibrosis Clinical Specimens by Targeting the cpn60 and 16S rRNA Genes","volume":"48","author":"Olson","year":"2010","journal-title":"J. Clin. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e0181721","DOI":"10.1128\/spectrum.01817-21","article-title":"Sequencing Independent Molecular Typing of Staphylococcus aureus Isolates: Approach for Infection Control and Clonal Characterization","volume":"10","author":"Dufkova","year":"2022","journal-title":"Microbiol. Spectr."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1043","DOI":"10.1177\/0194599819866692","article-title":"The Potential of High-Throughput DNA Sequencing of the Paranasal Sinus Microbiome in Diagnosing Odontogenic Sinusitis","volume":"161","author":"Haider","year":"2019","journal-title":"Otolaryngol. Head Neck Surg."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Wu, J., Zheng, M., Zhao, Y., Yin, W., Sima, Y., Zhao, J., Wang, X., Lin, J., and Zhang, L. (2023). Bacterial Diversity and Community Characteristics of the Sinus and Dental Regions in Adults with Odontogenic Sinusitis. BMC Microbiol., 23.","DOI":"10.1186\/s12866-023-02917-7"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1016\/j.jds.2015.06.001","article-title":"Pyrosequencing analysis of apical microbiota of teeth with persistent apical periodontitis","volume":"10","author":"Ping","year":"2015","journal-title":"J. Dent. Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1038\/s41368-024-00278-z","article-title":"Expert consensus on odontogenic maxillary sinusitis multi-disciplinary treatment","volume":"16","author":"Lin","year":"2024","journal-title":"Int. J. Oral Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1002\/alr.22627","article-title":"Comparison of bacterial maxillary sinus cultures between odontogenic sinusitis and chronic rhinosinusitis","volume":"11","author":"Bhargava","year":"2021","journal-title":"Int. Forum Allergy Rhinol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"134","DOI":"10.3390\/oral3010012","article-title":"Microbiology of Maxillary Sinus Infections: Systematic Review on the Relationship of Infectious Sinus Pathology with Oral Pathology","volume":"3","author":"Montero","year":"2023","journal-title":"Oral"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1002\/alr.21629","article-title":"Odontogenic and rhinogenic chronic sinusitis: A modern microbiological comparison","volume":"6","author":"Saibene","year":"2016","journal-title":"Int. Forum Allergy Rhinol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1","DOI":"10.4193\/Rhin20.401","article-title":"European Position Paper on Rhinosinusitis and Nasal Polyps 2020","volume":"58","author":"Fokkens","year":"2020","journal-title":"Rhinology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"216","DOI":"10.1016\/j.wjorl.2018.08.004","article-title":"The microbiome and chronic rhinosinusitis","volume":"4","author":"Sivasubramaniam","year":"2018","journal-title":"World J. Otorhinolaryngol. Head Neck Surg."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Michalik, M., and Krawczyk, B. (2024). Chronic Rhinosinusitis\u2014Microbiological Etiology, Potential Genetic Markers, and Diagnosis. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25063201"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"375","DOI":"10.5114\/ada.2018.77667","article-title":"Bacteriological analysis of isolated chronic sinusitis without polyps","volume":"35","author":"Janik","year":"2018","journal-title":"Post\u0119py Dermatol. Alergol."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Kim, D., Assiri, A.M., and Kim, J.H. (2019). Recent trends in bacteriology of adult patients with chronic rhinosinusitis. J. Clin. Med., 8.","DOI":"10.3390\/jcm8111889"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"823","DOI":"10.1097\/01.MLG.0000157332.17291.FC","article-title":"Microbiology of acute and chronic maxillary sinusitis associated with an odontogenic origin","volume":"115","author":"Brook","year":"2005","journal-title":"Laryngoscope"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1556\/1886.2020.00010","article-title":"The incidence of anaerobic bacteria in adult patients with chronic sinusitis: A prospective, single-centre microbiological study","volume":"10","author":"Torkos","year":"2020","journal-title":"Eur. J. Microbiol. Immunol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Albu, S., and Roman, A. (2025). Navigating the Treatment Landscape of Odontogenic Sinusitis: Current Trends and Future Directions. Medicina, 61.","DOI":"10.3390\/medicina61122175"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"21023","DOI":"10.1038\/s41598-022-24921-z","article-title":"Microbiota dysbiosis in odontogenic rhinosinusitis and its association with anaerobic bacteria","volume":"12","author":"Lu","year":"2022","journal-title":"Sci. Rep."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"6602","DOI":"10.1038\/srep06602","article-title":"Intraindividual variation in core microbiota in peri-implantitis and periodontitis","volume":"4","author":"Maruyama","year":"2014","journal-title":"Sci. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2009","DOI":"10.3402\/jom.v1i0.2009","article-title":"Distinctive features of the microbiota associated with different forms of apical periodontitis","volume":"1","author":"Siqueira","year":"2009","journal-title":"J. Oral Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Qian, W., Ma, T., Ye, M., Li, Z., Liu, Y., and Hao, P. (2019). Microbiota in the apical root canal system of tooth with apical periodontitis. BMC Genom., 20.","DOI":"10.1186\/s12864-019-5474-y"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Mussano, F., Ferrocino, I., Gavrilova, N., Genova, T., Dell\u2019Acqua, A., Cocolin, L., and Carossa, S. (2018). Apical periodontitis: Preliminary assessment of microbiota by 16S rRNA high throughput amplicon target sequencing. BMC Oral Health, 18.","DOI":"10.1186\/s12903-018-0520-8"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3130607","DOI":"10.1155\/2018\/3130607","article-title":"Composition Analysis and Feature Selection of the Oral Microbiota Associated with Periodontal Disease","volume":"2018","author":"Chen","year":"2018","journal-title":"Biomed Res. Int."},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Bordagaray, M.J., Fern\u00e1ndez, A., Garrido, M., Astorga, J., Hoare, A., and Hern\u00e1ndez, M. (2021). Systemic and Extraradicular Bacterial Translocation in Apical Periodontitis. Front. Cell. Infect. Microbiol., 11.","DOI":"10.3389\/fcimb.2021.649925"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"102925","DOI":"10.1016\/j.amjoto.2021.102925","article-title":"Diagnosing odontogenic sinusitis of endodontic origin: A multidisciplinary literature review","volume":"42","author":"Craig","year":"2021","journal-title":"Am. J. Otolaryngol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"156","DOI":"10.1038\/s41579-018-0129-6","article-title":"Fusobacterium nucleatum\u2014Symbiont, opportunist and oncobacterium","volume":"17","author":"Brennan","year":"2019","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Brooks, J.P., Edwards, D.J., Harwich, M.D., Rivera, M.C., Fettweis, J.M., Serrano, M.G., Reris, R.A., Sheth, N.U., Huang, B., and Girerd, P. (2015). The truth about metagenomics: Quantifying and counteracting bias in 16S rRNA studies Ecological and evolutionary microbiology. BMC Microbiol., 15.","DOI":"10.1186\/s12866-015-0351-6"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1016","DOI":"10.1038\/ismej.2012.174","article-title":"The subgingival microbiome in health and periodontitis and its relationship with community biomass and inflammation","volume":"7","author":"Abusleme","year":"2013","journal-title":"ISME J."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"259","DOI":"10.1038\/nrmicro.2017.14","article-title":"The microbiota of the respiratory tract: Gatekeeper to respiratory health","volume":"15","author":"Man","year":"2017","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Bouillaguet, S., Manoil, D., Girard, M., Louis, J., Ga\u00efa, N., Leo, S., Schrenzel, J., and Lazarevic, V. (2018). Root microbiota in primary and secondary apical periodontitis. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.02374"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"151ra124","DOI":"10.1126\/scitranslmed.3003783","article-title":"Sinus microbiome diversity depletion and Corynebacterium tuberculostearicum enrichment mediates rhinosinusitis","volume":"4","author":"Abreu","year":"2012","journal-title":"Sci. Transl. Med."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1002\/lio2.765","article-title":"Relationship between infected tooth extraction and improvement of odontogenic maxillary sinusitis","volume":"7","author":"Yoshida","year":"2022","journal-title":"Laryngoscope Investig. Otolaryngol."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"e00187-18","DOI":"10.1128\/msystems.00187-18","article-title":"New Insights into Human Nostril Microbiome from the Expanded Human Oral Microbiome Database (eHOMD): A Resource for the Microbiome of the Human Aerodigestive Tract","volume":"3","author":"Escapa","year":"2018","journal-title":"mSystems"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"e0007722","DOI":"10.1128\/msphere.00077-22","article-title":"Comparative Analysis and Data Provenance for 1113 Bacterial Genome Assemblies","volume":"7","author":"Yarmosh","year":"2022","journal-title":"mSphere"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"D733","DOI":"10.1093\/nar\/gkv1189","article-title":"Reference sequence (RefSeq) database at NCBI: Current status, taxonomic expansion, and functional annotation","volume":"44","author":"Wright","year":"2016","journal-title":"Nucleic Acids Res."},{"key":"ref_51","first-page":"D84","article-title":"GenBank","volume":"48","author":"Sayers","year":"2020","journal-title":"Nucleic Acids Res."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"W521","DOI":"10.1093\/nar\/gkae241","article-title":"The EMBL-EBI Job Dispatcher sequence analysis tools framework in 2024","volume":"52","author":"Madeira","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1189","DOI":"10.1093\/bioinformatics\/btp033","article-title":"Jalview Version 2\u2014A multiple sequence alignment editor and analysis workbench","volume":"25","author":"Waterhouse","year":"2009","journal-title":"Bioinformatics"},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Ye, J., Coulouris, G., Zaretskaya, I., Cutcutache, I., Rozen, S., and Madden, T.L. (2012). Primer-BLAST: A tool to design target-specific primers for polymerase chain reaction. BMC Bioinform., 13.","DOI":"10.1186\/1471-2105-13-134"},{"key":"ref_55","first-page":"127","article-title":"Nuclear DNA Content and Genome Size of Trout and Human","volume":"51","author":"Dolezel","year":"2023","journal-title":"Cytom. Part A"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"852","DOI":"10.1038\/s41587-019-0209-9","article-title":"Reproducible, interactive, scalable and extensible microbiome data science using QIIME 2","volume":"37","author":"Bolyen","year":"2019","journal-title":"Nat. Biotechnol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1186\/s40168-018-0470-z","article-title":"Optimizing taxonomic classification of marker-gene amplicon sequences with QIIME 2\u2032s q2-feature-classifier plugin","volume":"6","author":"Bokulich","year":"2018","journal-title":"Microbiome"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"D590","DOI":"10.1093\/nar\/gks1219","article-title":"The SILVA ribosomal RNA gene database project: Improved data processing and web-based tools","volume":"41","author":"Quast","year":"2013","journal-title":"Nucleic Acids Res."},{"key":"ref_59","unstructured":"Posit Team (2024). RStudio: Integrated Development Environment for, R, PBC. Available online: http:\/\/www.posit.co\/."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/9\/4010\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,30]],"date-time":"2026-04-30T10:48:06Z","timestamp":1777546086000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/9\/4010"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,30]]},"references-count":59,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2026,5]]}},"alternative-id":["ijms27094010"],"URL":"https:\/\/doi.org\/10.3390\/ijms27094010","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,30]]}}}