{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T09:44:05Z","timestamp":1785923045103,"version":"3.56.0"},"reference-count":91,"publisher":"MDPI AG","issue":"15","license":[{"start":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T00:00:00Z","timestamp":1785888000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003093","name":"Ministry of Higher Education","doi-asserted-by":"publisher","award":["FRGS\/1\/2022\/WAB04\/UPM\/02\/6"],"award-info":[{"award-number":["FRGS\/1\/2022\/WAB04\/UPM\/02\/6"]}],"id":[{"id":"10.13039\/501100003093","id-type":"DOI","asserted-by":"publisher"}]},{"award":["FRGS\/1\/2022\/WAB04\/UPM\/02\/6"],"award-info":[{"award-number":["FRGS\/1\/2022\/WAB04\/UPM\/02\/6"]}],"id":[{"id":"https:\/\/ror.org\/05mcs2t73","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biology"],"abstract":"<jats:p>Fish pathogens pose significant challenges to aquaculture, causing high mortality and economic losses. Thus, rapid, accurate, and cost-effective identification methods for disease management are needed. Conventional bacterial identification techniques are time-consuming and costly. Raman spectroscopy is an emerging alternative, enabling species differentiation based on molecular signatures. This study investigates surface-enhanced Raman spectroscopy (SERS) combined with principal component analysis with linear discriminant analysis (PCA-LDA) and partial least-squares discriminant analysis (PLS-DA) to classify three relevant Vibrio species: Vibrio alginolyticus, Vibrio parahaemolyticus, and Vibrio vulnificus. SERS spectra from bacterial cultures were analyzed to identify species-specific variations. Distinct clustering was observed, with V. vulnificus showing stronger amide-related signals, V. parahaemolyticus exhibiting higher sulfur-related Raman peaks, and V. alginolyticus showing distinct lipid region signals. The PCA-LDA model achieved 100% classification and 97.06% cross-validation accuracy under standardized laboratory conditions in this controlled proof-of-concept study using spectral replicates obtained from cultured laboratory bacterial stocks. The PLS-DA model showed excellent discriminatory performance (AUC = 1.00), with strong calibration for two species (R2 &gt; 0.80) and slightly lower for V. alginolyticus (R2 = 0.71). These findings demonstrate that SERS combined with multivariate analysis is a promising approach for rapid and label-free Vibrio identification and classification under controlled laboratory conditions.<\/jats:p>","DOI":"10.3390\/biology15151300","type":"journal-article","created":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T08:14:46Z","timestamp":1785917686000},"page":"1300","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Discrimination of Closely Related Vibrio Strains by Label-Free Surface-Enhanced Raman Spectroscopy (SERS)"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-6143-2253","authenticated-orcid":false,"given":"Aini Nadia","family":"Mazlan","sequence":"first","affiliation":[{"name":"Institute of Bioscience, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nathaniel","family":"Leong","sequence":"additional","affiliation":[{"name":"Wireless and Photonics Networks, Research Centre, Faculty of Engineering, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8577-386X","authenticated-orcid":false,"given":"Annie","family":"Christianus","sequence":"additional","affiliation":[{"name":"Department of Aquaculture, Faculty of Agriculture, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9385-1925","authenticated-orcid":false,"given":"Zuraidah","family":"Zan","sequence":"additional","affiliation":[{"name":"Department of Computer and Communication Systems, Faculty of Engineering, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Norazrin","family":"Ariffin","sequence":"additional","affiliation":[{"name":"Department of Agriculture Technology, Faculty of Agriculture, Universiti Putra Malaysia, Serdang 43400, Selangor, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0669-5339","authenticated-orcid":false,"given":"Junfeng","family":"Xie","sequence":"additional","affiliation":[{"name":"State Key Laboratory of Biocontrol, China-ASEAN Belt and Road Joint Laboratory on Mariculture Technology, School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3766-4372","authenticated-orcid":false,"given":"Annita Seok Kian","family":"Yong","sequence":"additional","affiliation":[{"name":"Institut Marin Borneo, Universiti Malaysia Sabah, Jalan UMS, Kota Kinabalu 88400, Sabah, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chou Min","family":"Chong","sequence":"additional","affiliation":[{"name":"Laboratory of Sustainable Aquaculture (AquaLab), International Institute of Aquaculture and Aquatic Sciences (I-AQUAS), Universiti Putra Malaysia, Port Dickson 71050, Negeri Sembilan, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"101459","DOI":"10.1016\/j.aqrep.2022.101459","article-title":"Pathology and Pathogenesis of Vibrio Infection in Fish: A Review","volume":"28","author":"Manchanayake","year":"2023","journal-title":"Aquac. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1002\/aah.10055","article-title":"Natural Concurrent Infection of Vibrio harveyi and V. alginolyticus in Cultured Hybrid Groupers in Malaysia","volume":"31","author":"Mohamad","year":"2019","journal-title":"J. Aquat. Anim. Health"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"e13051","DOI":"10.1111\/jwas.13051","article-title":"Cutting-edge Technologies for Detecting and Controlling Fish Diseases: Current Status, Outlook, and Challenges","volume":"55","author":"Islam","year":"2024","journal-title":"J. World Aquac. Soc."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"36","DOI":"10.4314\/ovj.v6i1.6","article-title":"Isolation and Molecular Identification of Vibrio spp. by Sequencing of 16S rDNA from Seafood, Meat and Meat Products in Libya","volume":"6","author":"Azwai","year":"2016","journal-title":"Open Vet. J."},{"key":"ref_5","unstructured":"Adetunji, C.O., Egbuna, C., Ficai, A., and Ijabadeniyi, O.A. (2024). Isolation Techniques Used for Molecular Characterization of Beneficial Microorganisms: Cultural, Biochemical and Molecular Characterization. Handbook of Agricultural Biotechnology, Wiley."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"475","DOI":"10.1007\/s10068-021-00877-0","article-title":"Development of Enhanced Selective Media for Detection of Vibrio parahaemolyticus in Oysters","volume":"30","author":"Yoon","year":"2021","journal-title":"Food Sci. Biotechnol."},{"key":"ref_7","first-page":"1","article-title":"Vibrio Related Diseases in Aquaculture and Development of Rapid and Accurate Identification Methods","volume":"3","author":"Chatterjee","year":"2012","journal-title":"J. Mar. Sci. Res. Dev."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"24","DOI":"10.1016\/j.mimet.2015.01.014","article-title":"Differential Specificity of Selective Culture Media for Enumeration of Pathogenic Vibrios: Advantages and Limitations of Multi-Plating Methods","volume":"111","author":"Nigro","year":"2015","journal-title":"J. Microbiol. Methods"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1016\/j.vibspec.2016.06.008","article-title":"Raman Spectroscopic Identification of Single Bacterial Cells at Different Stages of Their Lifecycle","volume":"86","author":"Hlaing","year":"2016","journal-title":"Vib. Spectrosc."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"34","DOI":"10.61751\/bmbr\/4.2023.34","article-title":"Microbiological Diagnostics: From Traditional to Molecular Genetic Methods: A Literature Review","volume":"5","author":"Ivashko","year":"2023","journal-title":"Bmbr"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Tang, Y.-W., and Stratton, C.W. (2018). Interpretation and Relevance of Advanced Technique Results. Advanced Techniques in Diagnostic Microbiology, Springer International Publishing.","DOI":"10.1007\/978-3-319-95111-9_31"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"836","DOI":"10.1111\/raq.12870","article-title":"Emerging Technologies Revolutionising Disease Diagnosis and Monitoring in Aquatic Animal Health","volume":"16","author":"Bohara","year":"2024","journal-title":"Rev. Aquac."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"e14023","DOI":"10.1111\/jfd.14023","article-title":"Mucosal Epithelial Homeostasis: Reference Intervals for Skin, Gill Lamellae and Filament for Atlantic Salmon and Other Fish Species","volume":"48","author":"Merkin","year":"2025","journal-title":"J. Fish Dis."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Cai, L., Fang, G., Tang, J., Cheng, Q., and Han, X. (2022). Label-Free Surface-Enhanced Raman Spectroscopic Analysis of Proteins: Advances and Applications. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232213868"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Agnello, S. (2021). Raman and Micro-Raman Spectroscopy. Spectroscopy for Materials Characterization, Wiley.","DOI":"10.1002\/9781119698029"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1007\/s40828-020-00129-4","article-title":"Introduction to Vibrational Spectroscopies","volume":"7","author":"Campanella","year":"2021","journal-title":"ChemTexts"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"315","DOI":"10.1364\/AOP.9.000315","article-title":"Raman Spectroscopy: Techniques and Applications in the Life Sciences","volume":"9","author":"Shipp","year":"2017","journal-title":"Adv. Opt. Photon."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Qiu, Y., Kuang, C., Liu, X., and Tang, L. (2022). Single-Molecule Surface-Enhanced Raman Spectroscopy. Sensors, 22.","DOI":"10.3390\/s22134889"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4756","DOI":"10.1002\/anie.201205748","article-title":"Surface-Enhanced Raman Spectroscopy: Concepts and Chemical Applications","volume":"53","year":"2014","journal-title":"Angew. Chem. Int. Ed."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"338474","DOI":"10.1016\/j.aca.2021.338474","article-title":"Recent Developments on Gold Nanostructures for Surface Enhanced Raman Spectroscopy: Particle Shape, Substrates and Analytical Applications. A Review","volume":"1168","year":"2021","journal-title":"Anal. Chim. Acta"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1133","DOI":"10.1002\/jrs.1734","article-title":"Reference Database of Raman Spectra of Biological Molecules","volume":"38","author":"Vandenabeele","year":"2007","journal-title":"J. Raman Spectrosc."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Zhang, S., Qi, Y., Tan, S.P.H., Bi, R., and Olivo, M. (2023). Molecular Fingerprint Detection Using Raman and Infrared Spectroscopy Technologies for Cancer Detection: A Progress Review. Biosensors, 13.","DOI":"10.3390\/bios13050557"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"2630","DOI":"10.1021\/ac3021888","article-title":"In Situ Strain-Level Detection and Identification of Vibrio parahaemolyticus Using Surface-Enhanced Raman Spectroscopy","volume":"85","author":"Xu","year":"2013","journal-title":"Anal. Chem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.tifs.2022.07.012","article-title":"A Review: Research Progress of SERS-Based Sensors for Agricultural Applications","volume":"128","author":"Liu","year":"2022","journal-title":"Trends Food Sci. Technol."},{"key":"ref_25","first-page":"e202300067","article-title":"SERS Materials with Small-Molecule Sensitivity for Biological Diagnosis","volume":"4","author":"Xu","year":"2024","journal-title":"Anal. Sens."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1002\/cyto.a.20682","article-title":"Vibrational Spectroscopy\u2014A Powerful Tool for the Rapid Identification of Microbial Cells at the Single-cell Level","volume":"75A","author":"Harz","year":"2009","journal-title":"Cytom. Part A"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/j.jmii.2024.11.014","article-title":"Identification of Staphylococcus Aureus, Enterococcus Faecium, Klebsiella Pneumoniae, Pseudomonas Aeruginosa and Acinetobacter Baumannii from Raman Spectra by Artificial Intelligent Raman Detection and Identification System (AIRDIS) with Machine Learning","volume":"58","author":"Lin","year":"2025","journal-title":"J. Microbiol. Immunol. Infect."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Liu, Z., Xue, Y., Yang, C., Li, B., and Zhang, Y. (2023). Rapid Identification and Drug Resistance Screening of Respiratory Pathogens Based on Single-Cell Raman Spectroscopy. Front. Microbiol., 14.","DOI":"10.3389\/fmicb.2023.1065173"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"255","DOI":"10.1016\/S0167-7012(02)00127-6","article-title":"Identification of Medically Relevant Microorganisms by Vibrational Spectroscopy","volume":"51","author":"Maquelin","year":"2002","journal-title":"J. Microbiol. Methods"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1652","DOI":"10.1039\/D5AN00115C","article-title":"Raman Spectroscopy as a Comprehensive Tool for Profiling Endospore-Forming Bacteria","volume":"150","author":"Salbreiter","year":"2025","journal-title":"Analyst"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4274","DOI":"10.1529\/biophysj.103.038604","article-title":"Raman Spectroscopy Detects Biochemical Changes Due to Proliferation in Mammalian Cell Cultures","volume":"88","author":"Short","year":"2005","journal-title":"Biophys. J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1039\/B506413A","article-title":"Surface-Enhanced Raman Scattering for the Rapid Discrimination of Bacteria","volume":"132","author":"Jarvis","year":"2006","journal-title":"Faraday Discuss."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2101","DOI":"10.1007\/s44211-024-00645-0","article-title":"Comparative Study of Machine-and Deep-Learning Based Classification Algorithms for Biomedical Raman Spectroscopy (RS): Case Study of RS Based Pathogenic Microbe Identification","volume":"40","author":"Guo","year":"2024","journal-title":"Anal. Sci."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"619","DOI":"10.1002\/jrs.5827","article-title":"Surface-enhanced Raman Spectroscopy Using Uncoated Gold Nanoparticles for Bacteria Discrimination","volume":"51","author":"Akanny","year":"2020","journal-title":"J. Raman Spectrosc."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Gu, P., Feng, Y.-Z., Zhu, L., Kong, L.-Q., Zhang, X., Zhang, S., Li, S.-W., and Jia, G.-F. (2020). Unified Classification of Bacterial Colonies on Different Agar Media Based on Hyperspectral Imaging and Machine Learning. Molecules, 25.","DOI":"10.3390\/molecules25081797"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Wang, W., Liu, J., Guo, S., Liu, L., Yuan, Q., Guo, L., and Pan, S. (2021). Identification of Vibrio parahaemolyticus and Vibrio spp. Specific Outer Membrane Proteins by Reverse Vaccinology and Surface Proteome. Front. Microbiol., 11.","DOI":"10.3389\/fmicb.2020.625315"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Brugger, S.D., Baumberger, C., Jost, M., Jenni, W., Brugger, U., and M\u00fchlemann, K. (2012). Automated Counting of Bacterial Colony Forming Units on Agar Plates. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0033695"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1802392","DOI":"10.1002\/smll.201802392","article-title":"Picoanalysis of Drugs in Biofluids with Quantitative Label-Free Surface-Enhanced Raman Spectroscopy","volume":"14","author":"Turzhitsky","year":"2018","journal-title":"Small"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"742014","DOI":"10.1016\/j.aquaculture.2024.742014","article-title":"Discrimination of Hypoxia Tolerant and Intolerant Hybrid Groupers Using Surface-Enhanced Raman Spectroscopy of Epidermal Mucus","volume":"598","author":"Leong","year":"2025","journal-title":"Aquaculture"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1469","DOI":"10.1002\/jrs.1797","article-title":"Strategy to Improve the Reproducibility of Colloidal SERS","volume":"38","author":"Tantra","year":"2007","journal-title":"J. Raman Spectrosc."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Radzol, A.R.M., Lee, K.Y., Mansor, W., and Azman, A. (2014). Optimization of Savitzky-Golay Smoothing Filter for Salivary Surface Enhanced Raman Spectra of Non Structural Protein 1. Proceedings of the TENCON 2014\u20132014 IEEE Region 10 Conference, Bangkok, Thailand, 22\u201325 October 2014, IEEE.","DOI":"10.1109\/TENCON.2014.7022409"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"20290","DOI":"10.1039\/D4RA01735H","article-title":"Differentiation of Different Dibenzothiophene (DBT) Desulfurizing Bacteria via Surface-Enhanced Raman Spectroscopy (SERS)","volume":"14","author":"Anwer","year":"2024","journal-title":"RSC Adv."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1665","DOI":"10.1039\/c2an36478f","article-title":"Amide I Vibrational Mode Suppression in Surface (SERS) and Tip (TERS) Enhanced Raman Spectra of Protein Specimens","volume":"138","author":"Kurouski","year":"2013","journal-title":"Analyst"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"5513","DOI":"10.1128\/JB.182.19.5513-5520.2000","article-title":"Relation of Capsular Polysaccharide Production and Colonial Cell Organization to Colony Morphology in Vibrio parahaemolyticus","volume":"182","author":"McCarter","year":"2000","journal-title":"J. Bacteriol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"8538","DOI":"10.1021\/ac801426m","article-title":"In Situ Surface-Enhanced Raman Scattering Analysis of Biofilm","volume":"80","author":"Ivleva","year":"2008","journal-title":"Anal. Chem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"126298","DOI":"10.1016\/j.saa.2025.126298","article-title":"Portable SERS for Salivary-Based Detection of Oral Pre-Malignant Lesions and Carcinomas: A Step toward Clinical Implementation","volume":"339","author":"Kaushik","year":"2025","journal-title":"Spectrochim. Acta Part A Mol. Biomol. Spectrosc."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Vaitiek\u016bnait\u0117, D., and Snitka, V. (2021). Differentiation of Closely Related Oak-Associated Gram-Negative Bacteria by Label-Free Surface Enhanced Raman Spectroscopy (SERS). Microorganisms, 9.","DOI":"10.3390\/microorganisms9091969"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1271","DOI":"10.1007\/s11745-006-0163-4","article-title":"The Fatty Acid Composition of a Vibrio alginolyticus Associated with the Alga Cladophora coelothrix. Identification of the Novel 9-methyl-10-hexadecenoic Acid","volume":"32","author":"Carballeira","year":"1997","journal-title":"Lipids"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"4631","DOI":"10.1007\/s00216-016-9540-x","article-title":"The Biochemical Origins of the Surface-Enhanced Raman Spectra of Bacteria: A Metabolomics Profiling by SERS","volume":"408","author":"Premasiri","year":"2016","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"5019","DOI":"10.1007\/s00216-018-1153-0","article-title":"Strain-Level Typing and Identification of Bacteria\u2014A Novel Approach for SERS Active Plasmonic Nanostructures","volume":"410","author":"Witkowska","year":"2018","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"Varmuza, K., and Filzmoser, P. (2016). Introduction to Multivariate Statistical Analysis in Chemometrics, CRC Press.","DOI":"10.1201\/9781420059496"},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Lasalvia, M., Capozzi, V., and Perna, G. (2022). A Comparison of PCA-LDA and PLS-DA Techniques for Classification of Vibrational Spectra. Appl. Sci., 12.","DOI":"10.3390\/app12115345"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"987","DOI":"10.1177\/0003702818764672","article-title":"Surface-Enhanced Raman Scattering (SERS) in Microbiology: Illumination and Enhancement of the Microbial World","volume":"72","author":"Chisanga","year":"2018","journal-title":"Appl. Spectrosc."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"103278","DOI":"10.1016\/j.pdpdt.2023.103278","article-title":"Surface-Enhanced Raman Spectroscopy (SERS) for the Characterization of Supernatants of Bacterial Cultures of Bacterial Strains Causing Sinusitis","volume":"41","author":"Saleem","year":"2023","journal-title":"Photodiagnosis Photodyn. Ther."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1961","DOI":"10.4103\/0366-6999.211874","article-title":"Label-Free Detection for a DNA Methylation Assay Using Raman Spectroscopy","volume":"130","author":"Kim","year":"2017","journal-title":"Chin. Med. J."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1187","DOI":"10.1016\/j.saa.2010.12.079","article-title":"Raman Spectra of Amino Acids and Their Aqueous Solutions","volume":"78","author":"Zhu","year":"2011","journal-title":"Spectrochim. Acta Part A Mol. Biomol. Spectrosc."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1773","DOI":"10.1007\/s11694-017-9558-6","article-title":"Label-Free SERS Detection of Salmonella Typhimurium on DNA Aptamer Modified AgNR Substrates","volume":"11","author":"Chen","year":"2017","journal-title":"Food Meas."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"2348","DOI":"10.1002\/jrs.6204","article-title":"Raman Spectroscopy in Cell Biology and Microbiology","volume":"52","author":"Pezzotti","year":"2021","journal-title":"J. Raman Spectrosc."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"2874","DOI":"10.1039\/C6AN00306K","article-title":"The Origin of the Band at around 730 cm\u22121 in the SERS Spectra of Bacteria: A Stable Isotope Approach","volume":"141","author":"Kubryk","year":"2016","journal-title":"Analyst"},{"key":"ref_60","doi-asserted-by":"crossref","unstructured":"Bao, Q., Bo, X., Chen, L., Ren, Y., Wang, H., Kwok, L.-Y., and Liu, W. (2023). Comparative Analysis Using Raman Spectroscopy of the Cellular Constituents of Lacticaseibacillus Paracasei Zhang in a Normal and Viable but Nonculturable State. Microorganisms, 11.","DOI":"10.3390\/microorganisms11051266"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1080\/05704928.2014.923902","article-title":"Raman Spectroscopy of Biological Tissues","volume":"50","author":"Talari","year":"2015","journal-title":"Appl. Spectrosc. Rev."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"52878","DOI":"10.1021\/acsami.4c10187","article-title":"Effect of Encapsulation of Lactobacillus casei in Alginate\u2013Tapioca Flour Microspheres Coated with Different Biopolymers on the Viability of Probiotic Bacteria","volume":"16","author":"Michalczyk","year":"2024","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"28270","DOI":"10.1021\/acs.analchem.5c05031","article-title":"High-Wavenumber Raman Spectroscopy for the Identification and Biochemical Characterization of Microbial Species","volume":"97","author":"Walter","year":"2025","journal-title":"Anal. Chem."},{"key":"ref_64","doi-asserted-by":"crossref","unstructured":"Pezzotti, G., Adachi, T., Kitagawa, K., Ikegami, S., Imamura, H., Yamamoto, T., Okuma, K., Matsuo, Y., Zhu, W., and Yasukochi, Y. (2026). Raman Signatures of Cnm-Positive Streptococcus Mutans: I, the Molecular Origin of Cerebral Microbleeds. Front. Microbiol., 17.","DOI":"10.3389\/fmicb.2026.1784125"},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1111\/j.1574-6968.2010.02139.x","article-title":"Monitoring and Rapid Quantification of Total Carotenoids in Rhodotorula Glutinis Cells Using Laser Tweezers Raman Spectroscopy: Monitoring of Carotenoids in R. glutinis Cells","volume":"314","author":"Tao","year":"2011","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_66","doi-asserted-by":"crossref","unstructured":"Wei, C., Li, M., and Zhao, X. (2018). Surface-Enhanced Raman Scattering (SERS) with Silver Nano Substrates Synthesized by Microwave for Rapid Detection of Foodborne Pathogens. Front. Microbiol., 9.","DOI":"10.3389\/fmicb.2018.02857"},{"key":"ref_67","doi-asserted-by":"crossref","unstructured":"Verde, A., Mangini, M., Manag\u00f2, S., Tramontano, C., Rea, I., Boraschi, D., Italiani, P., and De Luca, A.C. (2021). SERS Sensing of Bacterial Endotoxin on Gold Nanoparticles. Front. Immunol., 12.","DOI":"10.3389\/fimmu.2021.758410"},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"2001739","DOI":"10.1002\/advs.202001739","article-title":"Bacteria Detection: From Powerful SERS to Its Advanced Compatible Techniques","volume":"7","author":"Zhou","year":"2020","journal-title":"Adv. Sci."},{"key":"ref_69","doi-asserted-by":"crossref","unstructured":"Hassan, M., Zhao, Y., and Zughaier, S.M. (2024). Recent Advances in Bacterial Detection Using Surface-Enhanced Raman Scattering. Biosensors, 14.","DOI":"10.3390\/bios14080375"},{"key":"ref_70","doi-asserted-by":"crossref","first-page":"4862","DOI":"10.1039\/D4AN00854E","article-title":"Characterization of Vaginal Lactobacillus in Biologically Relevant Fluid Using Surface-Enhanced Raman Spectroscopy","volume":"149","author":"Locke","year":"2024","journal-title":"Analyst"},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"119912","DOI":"10.1016\/j.saa.2021.119912","article-title":"Surface Enhanced Raman Spectroscopic (SERS) Behavior of Phenylpyruvates Used in Heterogeneous Catalytic Asymmetric Cascade Reaction","volume":"260","author":"Firkala","year":"2021","journal-title":"Spectrochim. Acta Part A Mol. Biomol. Spectrosc."},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"107446","DOI":"10.1016\/j.mimet.2026.107446","article-title":"Exploring the Role of Phage Resistance by Using Label-Free SERS for Biochemical Profiling of Gram-Negative Pathogen Pseudomonas Aeruginosa Strains with Machine Learning Approach","volume":"244","author":"Zulfiqar","year":"2026","journal-title":"J. Microbiol. Methods"},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"3238","DOI":"10.1364\/BOE.5.003238","article-title":"Evaluation of Antibiotic Effects on Pseudomonas Aeruginosa Biofilm Using Raman Spectroscopy and Multivariate Analysis","volume":"5","author":"Jung","year":"2014","journal-title":"Biomed. Opt. Express"},{"key":"ref_74","doi-asserted-by":"crossref","first-page":"1343","DOI":"10.3390\/s7081343","article-title":"Raman Spectroscopy Cell-Based Biosensors","volume":"7","author":"Notingher","year":"2007","journal-title":"Sensors"},{"key":"ref_75","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1021\/ac034689c","article-title":"Discrimination of Bacteria Using Surface-Enhanced Raman Spectroscopy","volume":"76","author":"Jarvis","year":"2004","journal-title":"Anal. Chem."},{"key":"ref_76","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/1348-0421.12465","article-title":"Regulation Systems of Protease and Hemolysin Production in Vibrio vulnificus","volume":"61","author":"Elgaml","year":"2017","journal-title":"Microbiol. Immunol."},{"key":"ref_77","doi-asserted-by":"crossref","first-page":"5461","DOI":"10.1128\/IAI.71.10.5461-5471.2003","article-title":"Characterization and Pathogenic Significance of Vibrio vulnificus Antigens Preferentially Expressed in Septicemic Patients","volume":"71","author":"Kim","year":"2003","journal-title":"Infect. Immun."},{"key":"ref_78","doi-asserted-by":"crossref","first-page":"1053","DOI":"10.1007\/s00253-011-3634-3","article-title":"Rapid Detection of Food- and Waterborne Bacteria Using Surface-Enhanced Raman Spectroscopy Coupled with Silver Nanosubstrates","volume":"92","author":"Fan","year":"2011","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_79","doi-asserted-by":"crossref","first-page":"1356","DOI":"10.1039\/C5AN02175H","article-title":"Label-Free Mapping of Single Bacterial Cells Using Surface-Enhanced Raman Spectroscopy","volume":"141","author":"Wang","year":"2016","journal-title":"Analyst"},{"key":"ref_80","doi-asserted-by":"crossref","first-page":"11593","DOI":"10.1039\/D1NR00708D","article-title":"Surface-Enhanced Raman Spectroscopy for Bioanalysis and Diagnosis","volume":"13","author":"Tahir","year":"2021","journal-title":"Nanoscale"},{"key":"ref_81","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/j.jare.2022.11.010","article-title":"Recent Advances in Surface Enhanced Raman Spectroscopy for Bacterial Pathogen Identifications","volume":"51","author":"Usman","year":"2023","journal-title":"J. Adv. Res."},{"key":"ref_82","doi-asserted-by":"crossref","first-page":"2785","DOI":"10.1021\/acs.analchem.0c03681","article-title":"Sensible Functional Linear Discriminant Analysis Effectively Discriminates Enhanced Raman Spectra of Mycobacterium Species","volume":"93","author":"Cheng","year":"2021","journal-title":"Anal. Chem."},{"key":"ref_83","doi-asserted-by":"crossref","first-page":"289","DOI":"10.1007\/s10103-020-03028-9","article-title":"Biochemical Characterization of Pathogenic Bacterial Species Using Raman Spectroscopy and Discrimination Model Based on Selected Spectral Features","volume":"36","author":"Pacheco","year":"2021","journal-title":"Lasers Med. Sci."},{"key":"ref_84","doi-asserted-by":"crossref","unstructured":"Rahman, M.H.-U., Sikder, R., Tripathi, M., Zahan, M., Ye, T., Gnimpieba, Z.E., Jasthi, B.K., Dalton, A.B., and Gadhamshetty, V. (2024). Machine Learning-Assisted Raman Spectroscopy and SERS for Bacterial Pathogen Detection: Clinical, Food Safety, and Environmental Applications. Chemosensors, 12.","DOI":"10.3390\/chemosensors12070140"},{"key":"ref_85","doi-asserted-by":"crossref","first-page":"3806","DOI":"10.1039\/D3AN00679D","article-title":"Identification of Bacteria in Mixed Infection from Urinary Tract of Patient\u2019s Samples Using Raman Analysis of Dried Droplets","volume":"148","author":"Ryabchykov","year":"2023","journal-title":"Analyst"},{"key":"ref_86","doi-asserted-by":"crossref","first-page":"103692","DOI":"10.1016\/j.vibspec.2024.103692","article-title":"Discriminant Analysis of a Mixture of Pathogenic Bacteria into Different Types and Proportions by Surface-Enhanced Raman Scattering Spectroscopy Combined with Chemometric Methods","volume":"132","author":"Zhao","year":"2024","journal-title":"Vib. Spectrosc."},{"key":"ref_87","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1021\/acsabm.1c01020","article-title":"Raman Studies on Surface-Imprinted Polymers to Distinguish the Polymer Surface, Imprints, and Different Bacteria","volume":"5","author":"Thier","year":"2022","journal-title":"ACS Appl. Bio Mater."},{"key":"ref_88","doi-asserted-by":"crossref","first-page":"3165","DOI":"10.1007\/s00216-019-01784-4","article-title":"A Micro-Raman and Chemometric Study of Urinary Tract Infection-Causing Bacterial Pathogens in Mixed Cultures","volume":"411","author":"Yogesha","year":"2019","journal-title":"Anal. Bioanal. Chem."},{"key":"ref_89","doi-asserted-by":"crossref","unstructured":"Karad\u017ei\u0107 Banjac, M., Kova\u010devi\u0107, S., and Podunavac-Kuzmanovi\u0107, S. (2024). Ongoing Multivariate Chemometric Approaches in Bioactive Compounds and Functional Properties of Foods\u2014A Systematic Review. Processes, 12.","DOI":"10.3390\/pr12030583"},{"key":"ref_90","doi-asserted-by":"crossref","first-page":"100033","DOI":"10.1016\/j.fochms.2021.100033","article-title":"Insights on the Role of Chemometrics and Vibrational Spectroscopy in Fruit Metabolite Analysis","volume":"3","author":"Sultanbawa","year":"2021","journal-title":"Food Chem. Mol. Sci."},{"key":"ref_91","doi-asserted-by":"crossref","unstructured":"Piazzon, M.C., Naya-Catal\u00e0, F., Sim\u00f3-Mirabet, P., Picard-S\u00e1nchez, A., Roig, F.J., Calduch-Giner, J.A., Sitj\u00e0-Bobadilla, A., and P\u00e9rez-S\u00e1nchez, J. (2019). Sex, Age, and Bacteria: How the Intestinal Microbiota Is Modulated in a Protandrous Hermaphrodite Fish. Front. Microbiol., 10.","DOI":"10.3389\/fmicb.2019.02512"}],"container-title":["Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-7737\/15\/15\/1300\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,5]],"date-time":"2026-08-05T08:49:54Z","timestamp":1785919794000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-7737\/15\/15\/1300"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,8,5]]},"references-count":91,"journal-issue":{"issue":"15","published-online":{"date-parts":[[2026,8]]}},"alternative-id":["biology15151300"],"URL":"https:\/\/doi.org\/10.3390\/biology15151300","relation":{},"ISSN":["2079-7737"],"issn-type":[{"value":"2079-7737","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,8,5]]}}}