{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,18]],"date-time":"2026-10-18T02:59:35Z","timestamp":1792292375568,"version":"4.4.0"},"reference-count":39,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2024,3,26]],"date-time":"2024-03-26T00:00:00Z","timestamp":1711411200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>Lactic acid bacteria are considered an inexhaustible source of bioactive compounds; indeed, products from their metabolism are known to have immunomodulatory and anti-inflammatory activity. Recently, we demonstrated that Cell-Free Supernatants (CFS) obtained from Lactobacillus (L.) acidophilus, Lactiplantibacillus (L.) plantarum, Lacticaseibacillus (L.) rhamnosus, and Limosilactobacillus (L.) reuteri can impair Candida pathogenic potential in an in vitro model of epithelial vaginal infection. This effect could be ascribed to a direct effect of living lactic acid bacteria on Candida virulence and to the production of metabolites that are able to impair fungal virulence. In the present work, stemming from these data, we deepened our knowledge of CFS from these four lactic acid bacteria by performing a metabolomic analysis to better characterize their composition. By using an untargeted metabolomic approach, we detected consistent differences in the metabolites produced by these four different lactic acid bacteria. Interestingly, L. rhamnosus and L. acidophilus showed the most peculiar metabolic profiles. Specifically, after a hierarchical clustering analysis, L. rhamnosus and L. acidophilus showed specific areas of significantly overexpressed metabolites that strongly differed from the same areas in other lactic acid bacteria. From the overexpressed compounds in these areas, inosine from L. rhamnosus returned with the best identification profile. This molecule has been described as having antioxidant, anti-inflammatory, anti-infective, and neuroprotective properties. The biological significance of its overproduction by L. rhamnosus might be important in its probiotic and\/or postbiotic activity.<\/jats:p>","DOI":"10.3390\/microorganisms12040662","type":"journal-article","created":{"date-parts":[[2024,3,26]],"date-time":"2024-03-26T13:16:41Z","timestamp":1711459001000},"page":"662","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["An Untargeted Metabolomic Analysis of Lacticaseibacillus (L.) rhamnosus, Lactobacillus (L.) acidophilus, Lactiplantibacillus (L.) plantarum and Limosilactobacillus (L.) reuteri Reveals an Upregulated Production of Inosine from L. rhamnosus"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1369-0955","authenticated-orcid":false,"given":"Luca","family":"Spaggiari","sequence":"first","affiliation":[{"name":"Clinical and Experimental Medicine PhD Program, University of Modena and Reggio Emilia, 41125 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalia","family":"Pedretti","sequence":"additional","affiliation":[{"name":"Department of Surgical, Medical, Dental and Morphological Sciences with Interest in Transplant, Oncological and Regenerative Medicine, University of Modena and Reggio Emilia, 41124 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"Ricchi","sequence":"additional","affiliation":[{"name":"Clinical and Experimental Medicine PhD Program, University of Modena and Reggio Emilia, 41125 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Diego","family":"Pinetti","sequence":"additional","affiliation":[{"name":"Centro Interdipartimentale Grandi Strumenti, University of Modena and Reggio Emilia, 41125 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppina","family":"Campisciano","sequence":"additional","affiliation":[{"name":"Institute for Maternal and Child Health-IRCCS, Burlo Garofolo, 34137 Trieste, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1611-0813","authenticated-orcid":false,"given":"Francesco","family":"De Seta","sequence":"additional","affiliation":[{"name":"Department of Obstetrics and Gynecology, IRCCS San Raffaele Scientific Institute, University Vita and Salute, 20132 Milan, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Manola","family":"Comar","sequence":"additional","affiliation":[{"name":"Institute for Maternal and Child Health-IRCCS, Burlo Garofolo, 34137 Trieste, Italy"},{"name":"Department of Medical Sciences, University of Trieste, 34129 Trieste, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samyr","family":"Kenno","sequence":"additional","affiliation":[{"name":"Department of Surgical, Medical, Dental and Morphological Sciences with Interest in Transplant, Oncological and Regenerative Medicine, University of Modena and Reggio Emilia, 41124 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Ardizzoni","sequence":"additional","affiliation":[{"name":"Department of Surgical, Medical, Dental and Morphological Sciences with Interest in Transplant, Oncological and Regenerative Medicine, University of Modena and Reggio Emilia, 41124 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5299-8197","authenticated-orcid":false,"given":"Eva","family":"Pericolini","sequence":"additional","affiliation":[{"name":"Department of Surgical, Medical, Dental and Morphological Sciences with Interest in Transplant, Oncological and Regenerative Medicine, University of Modena and Reggio Emilia, 41124 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,3,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Latif, A., Shehzad, A., Niazi, S., Zahid, A., Ashraf, W., Iqbal, M.W., Rehman, A., Riaz, T., Aadil, R.M., and Khan, I.M. (2023). Probiotics: Mechanism of Action, Health Benefits and Their Application in Food Industries. Front. Microbiol., 14.","DOI":"10.3389\/fmicb.2023.1216674"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"202","DOI":"10.3390\/dairy1030015","article-title":"Lactic Acid Bacteria: Food Safety and Human Health Applications","volume":"1","author":"Ayivi","year":"2020","journal-title":"Dairy"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Tang, C., and Lu, Z. (2019). Health Promoting Activities of Probiotics. J. Food Biochem., 43.","DOI":"10.1111\/jfbc.12944"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Huang, R., Wu, F., Zhou, Q., Wei, W., Yue, J., Xiao, B., and Luo, Z. (2022). Lactobacillus and Intestinal Diseases: Mechanisms of Action and Clinical Applications. Microbiol. Res., 260.","DOI":"10.1016\/j.micres.2022.127019"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1626","DOI":"10.1007\/s12602-023-10045-x","article-title":"The Current and Future Perspectives of Postbiotics","volume":"15","author":"Liang","year":"2023","journal-title":"Probiotics Antimicrob. Proteins"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2939","DOI":"10.1007\/s00253-016-7343-9","article-title":"Bacteriocins of Lactic Acid Bacteria: Extending the Family","volume":"100","author":"Mu","year":"2016","journal-title":"Appl. Microbiol. Biotechnol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"48","DOI":"10.4161\/gmic.22328","article-title":"Antimicrobials: Strategies for Targeting Obesity and Metabolic Health?","volume":"4","author":"Murphy","year":"2013","journal-title":"Gut Microbes"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2117508","DOI":"10.1080\/19490976.2022.2117508","article-title":"The Clinical Evidence for Postbiotics as Microbial Therapeutics","volume":"14","author":"Mosca","year":"2022","journal-title":"Gut Microbes"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"3192","DOI":"10.1038\/s41467-022-30661-5","article-title":"Lactobacillus rhamnosus Colonisation Antagonizes Candida albicans by Forcing Metabolic Adaptations That Compromise Pathogenicity","volume":"13","author":"Last","year":"2022","journal-title":"Nat. Commun."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Thoda, C., and Touraki, M. (2023). Immunomodulatory Properties of Probiotics and Their Derived Bioactive Compounds. Appl. Sci., 13.","DOI":"10.3390\/app13084726"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Abdul Hakim, B.N., Xuan, N.J., and Oslan, S.N.H. (2023). A Comprehensive Review of Bioactive Compounds from Lactic Acid Bacteria: Potential Functions as Functional Food in Dietetics and the Food Industry. Foods, 12.","DOI":"10.3390\/foods12152850"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Spaggiari, L., Sala, A., Ardizzoni, A., De Seta, F., Singh, D.K., Gacser, A., Blasi, E., and Pericolini, E. (2022). Lactobacillus acidophilus, L. plantarum, L. rhamnosus, and L. reuteri Cell-Free Supernatants Inhibit Candida parapsilosis Pathogenic Potential upon Infection of Vaginal Epithelial Cells Monolayer and in a Transwell Coculture System In Vitro. Microbiol. Spectr., 10.","DOI":"10.1128\/spectrum.02696-21"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2293","DOI":"10.1159\/000493831","article-title":"Inosine\u2014A Multifunctional Treatment for Complications of Neurologic Injury","volume":"49","author":"Doyle","year":"2018","journal-title":"Cell Physiol. Biochem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3590","DOI":"10.1021\/acs.analchem.8b05592","article-title":"Systematic Error Removal Using Random Forest for Normalizing Large-Scale Untargeted Lipidomics Data","volume":"91","author":"Fan","year":"2019","journal-title":"Anal. Chem."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Z\u00fcllig, T., Zandl-Lang, M., Tr\u00f6tzm\u00fcller, M., Hartler, J., Plecko, B., and K\u00f6feler, H.C. (2020). A Metabolomics Workflow for Analyzing Complex Biological Samples Using a Combined Method of Untargeted and Target-List Based Approaches. Metabolites, 10.","DOI":"10.3390\/metabo10090342"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"326809","DOI":"10.5402\/2012\/326809","article-title":"Comparison of the Effects of Adenosine, Inosine, and Their Combination as an Adjunct to Reperfusion in the Treatment of Acute Myocardial Infarction","volume":"2012","author":"Shafy","year":"2012","journal-title":"ISRN Cardiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2097","DOI":"10.1021\/es5002105","article-title":"Identifying Small Molecules via High Resolution Mass Spectrometry: Communicating Confidence","volume":"48","author":"Schymanski","year":"2014","journal-title":"Environ. Sci. Technol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"605","DOI":"10.1038\/s41575-019-0173-3","article-title":"Probiotics and Prebiotics in Intestinal Health and Disease: From Biology to the Clinic","volume":"16","author":"Sanders","year":"2019","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Di Gioia, D., and Biavati, B. (2018). Probiotics and Prebiotics in Animal Health and Food Safety, Springer International Publishing.","DOI":"10.1007\/978-3-319-71950-4"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/bs.afnr.2020.06.001","article-title":"Postbiotics and Paraprobiotics: A Review of Current Evidence and Emerging Trends","volume":"Volume 94","year":"2020","journal-title":"Advances in Food and Nutrition Research"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Mart\u00edn, R., and Langella, P. (2019). Emerging Health Concepts in the Probiotics Field: Streamlining the Definitions. Front. Microbiol., 10.","DOI":"10.3389\/fmicb.2019.01047"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"693412","DOI":"10.3389\/fnut.2021.693412","article-title":"Therapeutic and Improving Function of Lactobacilli in the Prevention and Treatment of Cardiovascular-Related Diseases: A Novel Perspective from Gut Microbiota","volume":"8","author":"Zhao","year":"2021","journal-title":"Front. Nutr."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Peng, H., Zhou, G., Yang, X.-M., Chen, G.-J., Chen, H.-B., Liao, Z.-L., Zhong, Q.-P., Wang, L., Fang, X., and Wang, J. (2022). Transcriptomic Analysis Revealed Antimicrobial Mechanisms of Lactobacillus rhamnosus SCB0119 against Escherichia coli and Staphylococcus aureus. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms232315159"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1016\/j.jff.2017.05.011","article-title":"Adhesion and Invasion of Listeria monocytogenes and Interaction with Lactobacillus rhamnosus GG after Habituation on Fresh-Cut Pear","volume":"34","author":"Iglesias","year":"2017","journal-title":"J. Funct. Foods"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Muyyarikkandy, M.S., and Amalaradjou, M. (2017). Lactobacillus Bulgaricus, Lactobacillus rhamnosus and Lactobacillus paracasei Attenuate Salmonella enteritidis, Salmonella Heidelberg and Salmonella typhimurium Colonization and Virulence Gene Expression In Vitro. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18112381"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1186\/1757-4749-1-14","article-title":"Salmonella enterica Serovar Typhimurium Adhesion and Cytotoxicity during Epithelial Cell Stress Is Reduced by Lactobacillus rhamnosus GG","volume":"1","author":"Burkholder","year":"2009","journal-title":"Gut Pathog."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1226","DOI":"10.4014\/jmb.2205.05041","article-title":"The Functional Roles of Lactobacillus acidophilus in Different Physiological and Pathological Processes","volume":"32","author":"Gao","year":"2022","journal-title":"J. Microbiol. Biotechnol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"518","DOI":"10.1111\/jam.12544","article-title":"In Vitro Evaluation of Antimicrobial Activity of Lactobacillus rhamnosus IMC 501\u00ae, Lactobacillus paracasei IMC 502\u00ae and SYNBIO\u00ae against Pathogens","volume":"117","author":"Coman","year":"2014","journal-title":"J. Appl. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.archoralbio.2016.12.014","article-title":"Inhibitory Effects of Lactobacillus rhamnosus and Lactobacillus casei on Candida Biofilm of Denture Surface","volume":"76","author":"Song","year":"2017","journal-title":"Arch. Oral Biol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1180","DOI":"10.1111\/jam.12772","article-title":"Probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 Exhibit Strong Antifungal Effects against Vulvovaginal Candidiasis-causing Candida glabrata Isolates","volume":"118","author":"Chew","year":"2015","journal-title":"J. Appl. Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1016\/j.gde.2014.07.008","article-title":"Inosine in DNA and RNA","volume":"26","author":"Alseth","year":"2014","journal-title":"Curr. Opin. Genet. Dev."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1093\/nar\/gky1163","article-title":"Inosine Induces Context-Dependent Recoding and Translational Stalling","volume":"47","author":"Licht","year":"2019","journal-title":"Nucleic Acids Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"29626","DOI":"10.1074\/jbc.273.45.29626","article-title":"Axon Outgrowth Is Regulated by an Intracellular Purine-Sensitive Mechanism in Retinal Ganglion Cells","volume":"273","author":"Benowitz","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"18320","DOI":"10.1073\/pnas.0605135103","article-title":"Mst3b, a Purine-Sensitive Ste20-like Protein Kinase, Regulates Axon Outgrowth","volume":"103","author":"Irwin","year":"2006","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"13486","DOI":"10.1073\/pnas.96.23.13486","article-title":"Inosine Stimulates Extensive Axon Collateral Growth in the Rat Corticospinal Tract after Injury","volume":"96","author":"Benowitz","year":"1999","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"4199","DOI":"10.1038\/srep04199","article-title":"Oral Administration of Inosine Produces Antidepressant-like Effects in Mice","volume":"4","author":"Muto","year":"2014","journal-title":"Sci. Rep."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1016\/j.tips.2004.01.006","article-title":"Immunomodulatory and Neuroprotective Effects of Inosine","volume":"25","author":"Sitkovsky","year":"2004","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1013","DOI":"10.4049\/jimmunol.164.2.1013","article-title":"Inosine Inhibits Inflammatory Cytokine Production by a Posttranscriptional Mechanism and Protects Against Endotoxin-Induced Shock","volume":"164","author":"Kuhel","year":"2000","journal-title":"J. Immunol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1213","DOI":"10.1164\/ajrccm.164.7.2101013","article-title":"Inosine Reduces Systemic Inflammation and Improves Survival in Septic Shock Induced by Cecal Ligation and Puncture","volume":"164","author":"Liaudet","year":"2001","journal-title":"Am. J. Respir. Crit. Care Med."}],"container-title":["Microorganisms"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-2607\/12\/4\/662\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:18:55Z","timestamp":1760105935000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-2607\/12\/4\/662"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,3,26]]},"references-count":39,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2024,4]]}},"alternative-id":["microorganisms12040662"],"URL":"https:\/\/doi.org\/10.3390\/microorganisms12040662","relation":{},"ISSN":["2076-2607"],"issn-type":[{"value":"2076-2607","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,3,26]]}}}