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While human milk oligosaccharides are established prebiotics in the neonatal gut, their impact on the vaginal niche remains largely unexplored. This study investigated the effects of pooled human milk oligosaccharides on the growth and physiology of vaginal (Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus iners) and gut-derived (Lactobacillus reuteri, Lactobacillus rhamnosus) commensals. Growth analyses revealed that human milk oligosaccharides significantly and selectively stimulated growth across all vaginal strains tested, whereas gut commensals exhibited variable or inhibited growth. Carbohydrate utilization assays and comparative genomics against Bifidobacterium infantis showed that Lactobacillus crispatus and Lactobacillus reuteri lack the canonical metabolic machinery to catabolize human milk oligosaccharides. Instead, nitrogen utilization assays identified a glucose-dependent pathway where human milk oligosaccharides are associated with the depletion of primary amines and amino acids in Lactobacillus crispatus supernatants. These results suggest that human milk oligosaccharides act as noncatabolic modulators of vaginal lactobacilli. Collectively, these in vitro findings may warrant investigation of human milk oligosaccharides as modulators of vaginal commensal physiology in more complex experimental systems.<\/jats:p>","DOI":"10.1021\/acsinfecdis.6c00250","type":"journal-article","created":{"date-parts":[[2026,8,10]],"date-time":"2026-08-10T19:01:59Z","timestamp":1786388519000},"page":"3080-3091","source":"Crossref","is-referenced-by-count":0,"title":["Human Milk Oligosaccharides\nModulate Nitrogen Utilization\nin\n                    <i>Lactobacillus crispatus<\/i>\n                    in a Glucose-Dependent\nManner"],"prefix":"10.1021","volume":"12","author":[{"given":"Julie A.","family":"Talbert","sequence":"first","affiliation":[{"name":"Vanderbilt University , , , ,","place":["Nashville, Tennessee, United States, 37240"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-8178-2027","authenticated-orcid":true,"given":"Thomas L.","family":"Kalmer","sequence":"additional","affiliation":[{"name":"Vanderbilt University , , , ,","place":["Nashville, Tennessee, United States, 37240"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5666-7232","authenticated-orcid":true,"given":"Allison S.","family":"Walker","sequence":"additional","affiliation":[{"name":"Vanderbilt University , , , ,","place":["Nashville, Tennessee, United States, 37240"]},{"name":"Vanderbilt University , , , ,","place":["Nashville, Tennessee, United States, 37232"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jennifer A.","family":"Gaddy","sequence":"additional","affiliation":[{"name":"Vanderbilt University Medical Center , , , ,","place":["Nashville, Tennessee, United States, 37232"]},{"name":"Vanderbilt University Medical Center , , , ,","place":["Nashville, Tennessee, United States, 37232"]},{"name":"Tennessee Valley Healthcare Systems , , , ,","place":["Nashville, Tennessee, United States, 37212"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5362-7235","authenticated-orcid":true,"given":"Steven D.","family":"Townsend","sequence":"additional","affiliation":[{"name":"Vanderbilt University , , , ,","place":["Nashville, Tennessee, United States, 37240"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,10]]},"reference":[{"issue":"3","key":"2026091109094133500_cit1","doi-asserted-by":"publisher","first-page":"180","DOI":"10.1016\/j.imj.2022.08.004","article-title":"Correlation between human gut microbiome\nand diseases","volume":"1","author":"Madhogaria","year":"2022","journal-title":"Infect. Med."},{"issue":"1","key":"2026091109094133500_cit2","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1038\/s41392-022-00974-4","article-title":"Microbiota\nin health and diseases","volume":"7","author":"Hou","year":"2022","journal-title":"Signal Transduction\nTargeted Ther."},{"issue":"6","key":"2026091109094133500_cit3","doi-asserted-by":"publisher","first-page":"825","DOI":"10.1097\/MPG.0000000000000752","article-title":"Fecal microbiota\ncomposition of breast-fed infants\nis correlated with human milk oligosaccharides consumed","volume":"60","author":"Wang","year":"2015","journal-title":"J. Pediatr. Gastroenterol. Nutr."},{"issue":"1","key":"2026091109094133500_cit4","doi-asserted-by":"publisher","first-page":"15792","DOI":"10.1038\/s41598-020-72635-x","article-title":"Comparison\nof gut microbiota in exclusively breast-fed and formula-fed babies:\na study of 91 term infants","volume":"10","author":"Ma","year":"2020","journal-title":"Sci. Rep."},{"issue":"7","key":"2026091109094133500_cit5","doi-asserted-by":"publisher","first-page":"533","DOI":"10.3390\/pathogens13070533","article-title":"Difference in the Intestinal Microbiota between Breastfeed\nInfants and Infants Fed with Artificial Milk: A Systematic Review","volume":"13","author":"Inchingolo","year":"2024","journal-title":"Pathogens"},{"issue":"7","key":"2026091109094133500_cit6","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.0050177","article-title":"Development\nof the human infant intestinal microbiota","volume":"5","author":"Palmer","year":"2007","journal-title":"PLoS\nBiol."},{"issue":"7402","key":"2026091109094133500_cit7","doi-asserted-by":"publisher","first-page":"222","DOI":"10.1038\/nature11053","article-title":"Human gut\nmicrobiome viewed across age and geography","volume":"486","author":"Yatsunenko","year":"2012","journal-title":"Nature"},{"issue":"7285","key":"2026091109094133500_cit8","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1038\/nature08821","article-title":"A human\ngut microbial gene catalogue established by metagenomic sequencing","volume":"464","author":"Qin","year":"2010","journal-title":"Nature"},{"issue":"4","key":"2026091109094133500_cit9","doi-asserted-by":"publisher","first-page":"2782","DOI":"10.1099\/ijsem.0.004107","article-title":"A taxonomic note on the genus Lactobacillus:\nDescription of 23 novel genera, emended description of the genus Lactobacillus\nBeijerinck 1901, and union of Lactobacillaceae and Leuconostocaceae","volume":"70","author":"Zheng","year":"2020","journal-title":"Int. J. Syst. Evol. Microbiol."},{"key":"2026091109094133500_cit10","doi-asserted-by":"publisher","first-page":"1042189","DOI":"10.3389\/fphar.2022.1042189","article-title":"Gut microbiome and human health: Exploring how the probiotic genus\nLactobacillus modulate immune responses","volume":"13","author":"Rastogi","year":"2022","journal-title":"Front.\nPharmacol."},{"issue":"3\u20134","key":"2026091109094133500_cit11","doi-asserted-by":"publisher","first-page":"168","DOI":"10.1007\/s12602-011-9080-9","article-title":"Effect\nof Lactobacillus casei on the Production of\nPro-Inflammatory Markers in Streptozotocin-Induced Diabetic Rats","volume":"3","author":"Zarfeshani","year":"2011","journal-title":"Probiotics Antimicrob. Proteins"},{"issue":"1","key":"2026091109094133500_cit12","doi-asserted-by":"publisher","first-page":"2431643","DOI":"10.1080\/19490976.2024.2431643","article-title":"Probiotic\nsignificance of Lactobacillus strains: a comprehensive review on health\nimpacts, research gaps, and future prospects","volume":"16","author":"Shah","year":"2024","journal-title":"Gut Microbes"},{"issue":"supplement_1","key":"2026091109094133500_cit13","doi-asserted-by":"publisher","first-page":"4680","DOI":"10.1073\/pnas.1002611107","article-title":"Vaginal\nmicrobiome of reproductive-age women","volume":"108","author":"Ravel","year":"2011","journal-title":"Proc.\nNatl. Acad. Sci. 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Microbiol."},{"issue":"3","key":"2026091109094133500_cit19","doi-asserted-by":"publisher","first-page":"319","DOI":"10.1016\/S1286-4579(02)01544-7","article-title":"Use of Lactobacillus\nto prevent infection by pathogenic\nbacteria","volume":"4","author":"Reid","year":"2002","journal-title":"Microbes Infect."},{"issue":"6","key":"2026091109094133500_cit20","doi-asserted-by":"publisher","first-page":"0025","DOI":"10.1128\/microbiolspec.UTI-0025-2016","article-title":"The Vaginal\nMicrobiota and Urinary Tract Infection","volume":"4","author":"Stapleton","year":"2016","journal-title":"Microbiol.\nSpectrum"},{"issue":"6","key":"2026091109094133500_cit21","doi-asserted-by":"publisher","first-page":"687","DOI":"10.1097\/QAD.0000000000001768","article-title":"Vaginal\nmicrobiota and susceptibility to HIV","volume":"32","author":"Eastment","year":"2018","journal-title":"AIDS"},{"issue":"21","key":"2026091109094133500_cit22","doi-asserted-by":"publisher","first-page":"2039","DOI":"10.1056\/NEJMra1411426","article-title":"Pelvic inflammatory\ndisease","volume":"372","author":"Brunham","year":"2015","journal-title":"N. Engl. J. Med."},{"issue":"6","key":"2026091109094133500_cit23","doi-asserted-by":"publisher","first-page":"1012","DOI":"10.1038\/s41591-019-0450-2","article-title":"The vaginal microbiome and preterm\nbirth","volume":"25","author":"Fettweis","year":"2019","journal-title":"Nat. Med."},{"issue":"1","key":"2026091109094133500_cit24","doi-asserted-by":"publisher","first-page":"6","DOI":"10.1186\/s40168-016-0223-9","article-title":"The interaction between vaginal microbiota,\ncervical length, and vaginal progesterone treatment for preterm birth\nrisk","volume":"5","author":"Kindinger","year":"2017","journal-title":"Microbiome"},{"issue":"10","key":"2026091109094133500_cit25","doi-asserted-by":"publisher","first-page":"1869","DOI":"10.1007\/s10096-024-04915-7","article-title":"The association\nbetween lifestyle factors and the composition of the vaginal microbiota:\na review","volume":"43","author":"Morsli","year":"2024","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"2026091109094133500_cit26","doi-asserted-by":"publisher","first-page":"819958","DOI":"10.3389\/fmicb.2022.819958","article-title":"Healthy Vaginal Microbiota\nand Influence of Probiotics Across the Female Life Span","volume":"13","author":"Lehtoranta","year":"2022","journal-title":"Front. Microbiol."},{"issue":"6","key":"2026091109094133500_cit27","doi-asserted-by":"publisher","DOI":"10.1111\/aji.70026","article-title":"Vaginal Microbiome: Environmental,\nBiological, and Racial Influences on Gynecological Health Across the\nLifespan","volume":"92","author":"Dube\u0301-Zinatelli","year":"2024","journal-title":"Am. J. Reprod. Immunol."},{"issue":"9","key":"2026091109094133500_cit28","doi-asserted-by":"publisher","first-page":"1147","DOI":"10.1093\/glycob\/cws074","article-title":"Human milk\noligosaccharides: every baby needs a sugar mama","volume":"22","author":"Bode","year":"2012","journal-title":"Glycobiology"},{"key":"2026091109094133500_cit29","doi-asserted-by":"publisher","first-page":"104074","DOI":"10.1016\/j.jff.2020.104074","article-title":"Human milk oligosaccharides:\nShaping the infant gut microbiota and supporting health","volume":"72","author":"Walsh","year":"2020","journal-title":"J. Funct. Foods"},{"issue":"2","key":"2026091109094133500_cit30","doi-asserted-by":"publisher","first-page":"994","DOI":"10.1021\/acs.jafc.2c05734","article-title":"Effects of Human Milk\nOligosaccharides in Infant Health Based on Gut Microbiota Alteration","volume":"71","author":"Hu","year":"2023","journal-title":"J. Agric. Food Chem."},{"key":"2026091109094133500_cit31","doi-asserted-by":"publisher","first-page":"181","DOI":"10.3389\/fmed.2018.00181","article-title":"The Vaginal Microenvironment: The\nPhysiologic Role\nof Lactobacilli","volume":"5","author":"Amabebe","year":"2018","journal-title":"Front. Med."},{"issue":"4","key":"2026091109094133500_cit32","doi-asserted-by":"publisher","first-page":"624","DOI":"10.1097\/MPG.0000000000001520","article-title":"Effects of\nInfant Formula With Human Milk Oligosaccharides\non Growth and Morbidity: A Randomized Multicenter Trial","volume":"64","author":"Puccio","year":"2017","journal-title":"J. Pediatr. Gastroenterol. Nutr."},{"issue":"14","key":"2026091109094133500_cit33","doi-asserted-by":"publisher","first-page":"3087","DOI":"10.3390\/nu15143087","article-title":"Infant Formula Supplemented with Five Human Milk Oligosaccharides\nShifts the Fecal Microbiome of Formula-Fed Infants Closer to That\nof Breastfed Infants","volume":"15","author":"Holst","year":"2023","journal-title":"Nutrients"},{"issue":"12","key":"2026091109094133500_cit34","doi-asserted-by":"publisher","first-page":"BSR20191575","DOI":"10.1042\/bsr20191575","article-title":"The effect of Group B Streptococcus on maternal and infants\u2019\nprognosis in Guizhou, China","volume":"39","author":"Dai","year":"2019","journal-title":"Biosci. Rep."},{"key":"2026091109094133500_cit35","first-page":"0323","article-title":"Neonatal infections: Group B Streptococcus","volume":"2014","author":"Heath","year":"2014","journal-title":"BMJ Clin. Evid."},{"issue":"suppl_2","key":"2026091109094133500_cit36","doi-asserted-by":"publisher","first-page":"S133","DOI":"10.1093\/cid\/cix661","article-title":"Preterm Birth Associated\nWith Group B Streptococcus Maternal Colonization Worldwide: Systematic\nReview and Meta-analyses","volume":"65","author":"Bianchi-Jassir","year":"2017","journal-title":"Clin. Infect. Dis."},{"issue":"11","key":"2026091109094133500_cit37","doi-asserted-by":"publisher","first-page":"1437","DOI":"10.1111\/1471-0528.13527","article-title":"Maternal\nGroup B Streptococcus-related stillbirth:\na systematic review","volume":"122","author":"Nan","year":"2015","journal-title":"BJOG"},{"key":"2026091109094133500_cit38","unstructured":"Schrag, S.; Gorwitz, R.; Fultz-Butts, K.; Schuchat, A.\n          Prevention\nof\nperinatal group B streptococcal disease. Revised guidelines from CDC.  MMWR Recommendations and Reports: Morbidity and Mortality\nWeekly Report Recommendations and Reports; Centers for Disease Control and Prevention, 2002; 51(RR-11), pp 1\u201322."},{"issue":"2","key":"2026091109094133500_cit39","doi-asserted-by":"publisher","DOI":"10.1542\/peds.2019-1881","article-title":"Management of Infants at Risk for Group B Streptococcal\nDisease","volume":"144","author":"Puopolo","year":"2019","journal-title":"Pediatrics"},{"issue":"3","key":"2026091109094133500_cit40","doi-asserted-by":"publisher","first-page":"315","DOI":"10.1021\/acsinfecdis.7b00183","article-title":"Antimicrobial and\nAntibiofilm Activity\nof Human Milk Oligosaccharides against Streptococcus agalactiae, Staphylococcus\naureus, and Acinetobacter baumannii","volume":"4","author":"Ackerman","year":"2018","journal-title":"ACS Infect.\nDis."},{"issue":"8","key":"2026091109094133500_cit41","doi-asserted-by":"publisher","first-page":"595","DOI":"10.1021\/acsinfecdis.7b00064","article-title":"Human Milk Oligosaccharides\nExhibit Antimicrobial and\nAntibiofilm Properties against Group B Streptococcus","volume":"3","author":"Ackerman","year":"2017","journal-title":"ACS Infect. Dis."},{"issue":"9","key":"2026091109094133500_cit42","doi-asserted-by":"publisher","first-page":"1737","DOI":"10.1021\/acscentsci.3c00101","article-title":"The Utility\nof Human Milk Oligosaccharides\nagainst Group B Streptococcus Infections of Reproductive Tissues and\nCognate Adverse Pregnancy Outcomes","volume":"9","author":"Moore","year":"2023","journal-title":"ACS Cent.\nSci."},{"issue":"6","key":"2026091109094133500_cit43","doi-asserted-by":"publisher","DOI":"10.1002\/cbic.202200643","article-title":"Anti-biofilm Activity\nof Human Milk Oligosaccharides\nin Clinical Strains of Streptococcus agalactiae with Diverse Capsular\nand Sequence Types","volume":"24","author":"Moore","year":"2023","journal-title":"ChemBioChem"},{"issue":"6","key":"2026091109094133500_cit44","doi-asserted-by":"publisher","first-page":"1729","DOI":"10.1021\/acsinfecdis.5c00295","article-title":"Human Milk\nOligosaccharides Mediate the Host\u2013Microbe Interface in a Model\nVaginal Community","volume":"11","author":"Talbert","year":"2025","journal-title":"ACS Infect. Dis."},{"issue":"12","key":"2026091109094133500_cit45","doi-asserted-by":"publisher","first-page":"3254","DOI":"10.1021\/acsinfecdis.1c00420","article-title":"Antibiofilm\nActivity of Human Milk Oligosaccharides against Multidrug Resistant\nand Susceptible Isolates of Acinetobacter baumannii","volume":"7","author":"Spicer","year":"2021","journal-title":"ACS Infect. Dis."},{"issue":"1","key":"2026091109094133500_cit46","doi-asserted-by":"publisher","DOI":"10.1128\/msphere.00885-21","article-title":"Human Milk\nOligosaccharides Reduce Murine Group B Streptococcus Vaginal Colonization\nwith Minimal Impact on the Vaginal Microbiota","volume":"7","author":"Mejia","year":"2022","journal-title":"mSphere"},{"issue":"10","key":"2026091109094133500_cit47","doi-asserted-by":"publisher","first-page":"7825","DOI":"10.3168\/jds.2017-12753","article-title":"Human milk oligosaccharide\nconsumption by probiotic and human-associated bifidobacteria and lactobacilli","volume":"100","author":"Thongaram","year":"2017","journal-title":"J. Dairy Sci."},{"issue":"1","key":"2026091109094133500_cit48","doi-asserted-by":"publisher","first-page":"35045","DOI":"10.1038\/srep35045","article-title":"A novel gene cluster allows preferential utilization\nof fucosylated milk oligosaccharides in Bifidobacterium longum subsp.\nlongum SC596","volume":"6","author":"Garrido","year":"2016","journal-title":"Sci. Rep."},{"key":"2026091109094133500_cit49","doi-asserted-by":"publisher","first-page":"1893","DOI":"10.3390\/nu16121893","article-title":"Variation in the Conservation of\nSpecies-Specific Gene Sets for HMO Degradation and Its Effects on\nHMO Utilization in Bifidobacteria","volume":"16","author":"Hermes","year":"2024","journal-title":"Nutrients"},{"key":"2026091109094133500_cit50","doi-asserted-by":"publisher","first-page":"1040721","DOI":"10.3389\/fmolb.2023.1040721","article-title":"Structure of co-expression networks of Bifidobacterium\nspecies in response to human milk oligosaccharides","volume":"10","author":"Gonza\u0301lez-Morelo","year":"2023","journal-title":"Front. Mol. 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