{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,21]],"date-time":"2026-10-21T04:34:04Z","timestamp":1792557244268,"version":"4.5.0"},"reference-count":54,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2021,10,1]],"date-time":"2021-10-01T00:00:00Z","timestamp":1633046400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell. Infect. Microbiol."],"abstract":"\n Background<\/jats:title>\n Bacterial infections are usually suspected in infertile couples seeking IVF with no clear understanding of the microbial compositions present in the seminal fluids and vaginal niche of the patients. We used next-generation sequencing technology to correlate microbiota compositions with IVF clinical outcomes.<\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n Thirty-six couples were recruited to provide seminal fluids and vaginal swabs. Bacterial DNA was extracted, and V4 region of the 16S rRNA was amplified and sequenced in a pair-end configuration on the Illumina MiSeq platform rendering 2 \u00d7 150 bp sequences. Microbial taxonomy to species level was generated using the Greengenes database. Linear discriminant analysis (LDA) effect size (LEfSe) was used to identify biologically and statistically significant differences in relative abundance.<\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n Seminal fluid microbiota compositions had lower bacterial concentrations compared with the vagina, but species diversity was significantly higher in seminal fluid samples. Azoospermic subjects had more relative abundance of\n Mycoplasma<\/jats:italic>\n and\n Ureaplasma.<\/jats:italic>\n In Normospermic semen,\n Lactobacillus<\/jats:italic>\n (43.86%) was the most abundant, followed by\n Gardnerella<\/jats:italic>\n (25.45%), while the corresponding vaginal samples,\n Lactobacillus<\/jats:italic>\n (61.74%) was the most abundant, followed by\n Prevotella<\/jats:italic>\n (6.07%) and\n Gardnerella<\/jats:italic>\n (5.86%).\n <\/jats:p>\n <\/jats:sec>\n \n Conclusions<\/jats:title>\n \n Semen samples with positive IVF were significantly colonized by\n Lactobacillus jensenii<\/jats:italic>\n (\n P<\/jats:italic>\n =0.002),\n Faecalibacterium<\/jats:italic>\n (\n P<\/jats:italic>\n =0.042) and significantly less colonized by\n Proteobacteria<\/jats:italic>\n ,\n Prevotella<\/jats:italic>\n ,\n Bacteroides<\/jats:italic>\n , and lower\n Firmicutes<\/jats:italic>\n \/\n Bacteroidetes<\/jats:italic>\n ratio compared with semen samples with negative IVF. Vaginal samples with positive IVF clinical outcome were significantly colonized by\n Lactobacillus gasseri<\/jats:italic>\n , less colonized by\n Bacteroides<\/jats:italic>\n and\n Lactobacillus iners.<\/jats:italic>\n This study has opened a window of possibility for\n Lactobacillus<\/jats:italic>\n replenishments in men and women before IVF treatment.\n <\/jats:p>\n <\/jats:sec>","DOI":"10.3389\/fcimb.2021.709372","type":"journal-article","created":{"date-parts":[[2021,10,1]],"date-time":"2021-10-01T12:23:29Z","timestamp":1633091009000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":54,"title":["Microbiome Compositions From Infertile Couples Seeking In Vitro Fertilization, Using 16S rRNA Gene Sequencing Methods: Any Correlation to Clinical Outcomes?"],"prefix":"10.3389","volume":"11","author":[{"given":"Somadina I.","family":"Okwelogu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joseph I.","family":"Ikechebelu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nneka R.","family":"Agbakoba","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kingsley C.","family":"Anukam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,10,1]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1007\/s15010-017-1075-3","article-title":"Improvement of Semen Parameters After Antibiotic Therapy in Asymptomatic Infertile Men Infected With Mycoplasma Genitalium","volume":"46","author":"Ahmadi","year":"2018","journal-title":"Infection"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1186\/2193-1801-2-369","article-title":"Bias Problems in Culture-Independent Analysis of Environmental Bacterial Communities: A Representative Study on Hydrocarbonoclastic Bacteria","volume":"2","author":"Al-Awadhi","year":"2013","journal-title":"Springerplus"},{"key":"B3","doi-asserted-by":"publisher","first-page":"e0176555","DOI":"10.1371\/journal.pone.0176555","article-title":"16s rRNA Gene Sequencing and Healthy Reference Ranges for 28 Clinically Relevant Microbial Taxa From the Human Gut Microbiome","volume":"12","author":"Almonacid","year":"2017","journal-title":"PloS One"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.3389\/fmed.2018.00181","article-title":"The Vaginal Microenvironment: The Physiologic Role of Lactobacilli","volume":"5","author":"Amabebe","year":"2018","journal-title":"Front. Med."},{"key":"B5","doi-asserted-by":"publisher","first-page":"96\u2212104","DOI":"10.4103\/TJOG.TJOG_67_18","article-title":"Vaginal Bacteriome of Nigerian Women in Health and Disease: A Study With 16S rRNA Metagenomics","volume":"36","author":"Anukam","year":"2019","journal-title":"Trop. J. Obstet. Gynaecol."},{"key":"B6","doi-asserted-by":"publisher","first-page":"924","DOI":"10.1086\/428288","article-title":"Factors Associated With Absence of H2O2-Producing Lactobacillus Among Women With Bacterial Vaginosis","volume":"191","author":"Beigi","year":"2005","journal-title":"J. Infect. Dis."},{"key":"B7","doi-asserted-by":"publisher","first-page":"462","DOI":"10.1093\/humupd\/1.5.462","article-title":"Anaerobes in Ejaculates of Subfertile Men","volume":"1","author":"Eggert-Kruse","year":"1995","journal-title":"Hum. Reprod. Update"},{"key":"B8","first-page":"3805","article-title":"Common Bacterial Isolates Associated With Semen of Men Attending the Fertility Clinic of the University of Teaching Hospital (U.B.T.H) Benin City, Nigeria","volume":"5","author":"Ekhaise","year":"2001","journal-title":"Afr J. Microbiol."},{"key":"B9","doi-asserted-by":"publisher","first-page":"1364\u22121371","DOI":"10.1016\/j.fertnstert.2015.10.012","article-title":"Reproductive Tract Microbiome in Assisted Reproductive Technologies","volume":"104","author":"Franasiak","year":"2015","journal-title":"Fertil Steril"},{"key":"B10","doi-asserted-by":"publisher","first-page":"842652","DOI":"10.1155\/2011\/842652","article-title":"Association Between Semen Exposure and Incident Bacterial Vaginosis","volume":"2011","author":"Gallo","year":"2011","journal-title":"Infect. Dis. Obstet Gynecol"},{"key":"B11","doi-asserted-by":"publisher","first-page":"103","DOI":"10.1016\/j.rbmo.2017.04.005","article-title":"What Fertility Specialists Should Know About the Vaginal Microbiome: A Review","volume":"35","author":"Garcia-Velasco","year":"2017","journal-title":"Reprod. BioMed. Online"},{"key":"B12","doi-asserted-by":"publisher","first-page":"198","DOI":"10.2164\/jandrol.107.003566","article-title":"Assessment of Chlamydia Trachomatis, Ureaplasma Urealyticum, Ureaplasma Parvum, Mycoplasma Hominis, and Mycoplasma Genitalium in Semen and First Void Urine Specimens of Asymptomatic Male Partners of Infertile Couples","volume":"29","author":"Gdoura","year":"2008","journal-title":"J. Androl."},{"key":"B13","doi-asserted-by":"publisher","first-page":"379","DOI":"10.1007\/s00404-010-1726-4","article-title":"Chlamydia, Mycoplasma and Ureaplasma Infections in Infertile Couples and Effects of These Infections on Fertility","volume":"283","author":"G\u00fcnyeli","year":"2011","journal-title":"Arch. Gynecol. Obstet."},{"key":"B14","doi-asserted-by":"publisher","first-page":"1809","DOI":"10.1093\/infdis\/jiy744","article-title":"Vaginal Microbiota and In Vitro Fertilization Outcomes: Development of a Simple Diagnostic Tool to Predict Patients at Risk of a Poor Reproductive Outcome","volume":"219","author":"Haahr","year":"2019","journal-title":"J. Infect. Dis."},{"key":"B15","doi-asserted-by":"publisher","first-page":"178","DOI":"10.1016\/j.rbmo.2019.04.006","article-title":"Vaginal Microbiota and IVF Outcomes: Poor Diagnosis Results in Flawed Conclusions","volume":"39","author":"Haahr","year":"2019","journal-title":"Repro BioMed. Online"},{"key":"B16","doi-asserted-by":"publisher","first-page":"795","DOI":"10.1093\/humrep\/dew026","article-title":"Abnormal Vaginal Microbiota may be Associated With Poor Reproductive Outcomes: A Prospective Study in IVF Patients","volume":"31","author":"Haahr","year":"2016","journal-title":"Hum. Reprod. Oxf Engl."},{"key":"B17","doi-asserted-by":"publisher","first-page":"1737","DOI":"10.1056\/NEJM199512283332604","article-title":"Association Between Bacterial Vaginosis and Preterm Delivery of a Low-Birth-Weight Infant. The Vaginal Infections and Prematurity Study Group","volume":"333","author":"Hillier","year":"1995","journal-title":"N Engl. J. Med."},{"key":"B18","doi-asserted-by":"publisher","first-page":"1261","DOI":"10.1016\/j.fertnstert.2013.07.1991","article-title":"Microbiota of the Seminal Fluid From Healthy and Infertile Men","volume":"100","author":"Hou","year":"2013","journal-title":"Fertil Steril"},{"key":"B19","first-page":"65","article-title":"Bacteriospermia and Sperm Quality in Infertile Male Patient at University of Benin Teaching Hospital, Benin City, Nigeria","volume":"4","author":"Ibadin","year":"2008","journal-title":"Malaysian J. Microbiol."},{"key":"B20","first-page":"69","article-title":"Serodynamics of Chlamydia Trachomatis Infection in Women With In-Vitro Fertilization-Embryo Transfer Failures","volume":"1","author":"Ibadin","year":"2013","journal-title":"J. Infertl. Reprod. Biol."},{"key":"B21","first-page":"1","article-title":"Assisted Reproduction Techniques (ART): The State of Art in Nigeria","volume":"8","author":"Ikechebelu","year":"2003","journal-title":"J. Clin. Med."},{"key":"B22","doi-asserted-by":"publisher","first-page":"657","DOI":"10.1080\/01443610310001604475","article-title":"High Prevalence of Male Infertility in Southeastern, Nigeria","volume":"23","author":"Ikechebelu","year":"2003","journal-title":"J. Obstet. Gynaecol."},{"key":"B23","doi-asserted-by":"publisher","first-page":"2","DOI":"10.4103\/2468-8452.189960","article-title":"Outcome of in-Vitro Fertilization Procedure at a Private Fertility Center in Nnewi, South-East Nigeria","volume":"1","author":"Ikechebelu","year":"2016","journal-title":"Afr. J. Infert. Assisted Conception"},{"key":"B24","doi-asserted-by":"publisher","first-page":"462","DOI":"10.1111\/j.1365-2605.2008.00878.x","article-title":"Microflora of the Seminal Fluid of Healthy Men and Men Suffering From Chronic Prostatitis Syndrome","volume":"32","author":"Ivanov","year":"2009","journal-title":"Int. J. Androl."},{"key":"B25","doi-asserted-by":"publisher","first-page":"23027","DOI":"10.1038\/srep23027","article-title":"Discovery of a Novel Seminal Fluid Microbiome and Influence of Estrogen Receptor Alpha Genetic Status","volume":"6","author":"Javurek","year":"2016","journal-title":"Sci. Rep."},{"key":"B26","doi-asserted-by":"publisher","first-page":"D199","DOI":"10.1093\/nar\/gkt1076","article-title":"Data, Information, Knowledge and Principle: Back to Metabolism in KEGG","volume":"42","author":"Kanehisa","year":"2014","journal-title":"Nucleic Acids Res."},{"key":"B27","doi-asserted-by":"publisher","first-page":"e1002863","DOI":"10.1371\/journal.pcbi.1002863","article-title":"A Guide to Enterotypes Across the Human Body: Meta-Analysis of Microbial Community Structures in Human Microbiome Datasets","volume":"9","author":"Koren","year":"2013","journal-title":"PloS Comput. Biol."},{"key":"B28","doi-asserted-by":"publisher","first-page":"814","DOI":"10.1038\/nbt.2676","article-title":"Predictive Functional Profiling of Microbial Communities Using 16S rRNA Marker Gene Sequences","volume":"31","author":"Langille","year":"2013","journal-title":"Nat. Biotechnol."},{"key":"B29","doi-asserted-by":"publisher","first-page":"418","DOI":"10.1093\/cid\/civ312","article-title":"Mycoplasma Genitalium Infection and Female Reproductive Tract Disease: A Meta-Analysis","volume":"61","author":"Lis","year":"2015","journal-title":"Clin. Infect. Dis."},{"key":"B30","doi-asserted-by":"publisher","first-page":"440","DOI":"10.1016\/j.resmic.2015.03.009","article-title":"Complementary Seminovaginal Microbiome in Couples","volume":"166","author":"Mandar","year":"2015","journal-title":"Res. Microbiol."},{"key":"B31","first-page":"22","article-title":"Infertility in Male: Risk Factors, Causes and Management-A Review","volume":"9","author":"Momoh","year":"2007","journal-title":"Benin J. Post Grad Med."},{"key":"B32","doi-asserted-by":"publisher","first-page":"86","DOI":"10.5281\/zenodo.4007804","article-title":"16s rRNA Metagenomics of Seminal Fluids From Medical Microbiology Laboratory in a Tertiary Hospital, Southern Nigeria","volume":"29","author":"Ndiokwere","year":"2019","journal-title":"J. Med. Lab. Sci."},{"key":"B33","doi-asserted-by":"publisher","first-page":"e14116","DOI":"10.1371\/journal.pone.0014116","article-title":"Characteristic Male Urine Microbiomes Associate With Asymptomatic Sexually Transmitted Infection","volume":"5","author":"Nelson","year":"2010","journal-title":"PloS One"},{"key":"B34","first-page":"4","article-title":"Microbiologic Review of Seminal Fluids in a Nigerian Tertiary Health Centre","volume":"7","author":"Nwadioha","year":"2016","journal-title":"Arch. Clin. Microbiol."},{"key":"B35","doi-asserted-by":"publisher","first-page":"1","DOI":"10.5281\/zenodo.4010799","article-title":"Comparative Abundance and Functional Biomarkers of the Vaginal and Gut Microbiome of Nigerian Women With Bacterial Vaginosis: A Study With 16S rRNA Metagenomics","volume":"29","author":"Okoli","year":"2019","journal-title":"J. Med. Lab. Sci."},{"key":"B36","first-page":"210","article-title":"Studies on the Significance of Positive Bacterial Semen Cultures in Male Fertility in Nigeria","volume":"46","author":"Onemu","year":"2001","journal-title":"Int. J. Fertil Womens Med."},{"key":"B37","doi-asserted-by":"publisher","first-page":"392","DOI":"10.1099\/mic.0.034280-0","article-title":"Analysis of Adherence, Biofilm Formation and Cytotoxicity Suggests a Greater Virulence Potential of Gardnerella Vaginalis Relative to Other Bacterial-Vaginosis Associated Anaerobes","volume":"156","author":"Patterson","year":"2010","journal-title":"Microbiol"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2013.00090","article-title":"Iron, Copper, Zinc, and Manganese Transport and Regulation in Pathogenic Enterobacteria: Correlations Between Strains, Site of Infection and the Relative Importance of the Different Metal Transport Systems for Virulence","volume":"5","author":"Porcheron","year":"2013","journal-title":"Front. Cell Infect. Microbiol."},{"key":"B39","doi-asserted-by":"publisher","first-page":"D590","DOI":"10.1093\/nar\/gks1219","article-title":"The SILVA Ribosomal RNA Gene Database Project: Improved Data Processing and Webbased Tools","volume":"41","author":"Quast","year":"2013","journal-title":"Nucleic Acids Res."},{"key":"B40","doi-asserted-by":"publisher","first-page":"4680","DOI":"10.1073\/pnas.1002611107","article-title":"Vaginal Microbiome of Reproductive-Age Women","volume":"108","author":"Ravel","year":"2010","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"B41","doi-asserted-by":"publisher","first-page":"1977","DOI":"10.1128\/JCM.41.5.1977-1986.2003","article-title":"Microbial Population Diversity in the Urethras of Healthy Males and Males Suffering From non-Chlamydial, Nongonococcal Urethritis","volume":"41","author":"Riemersma","year":"2003","journal-title":"J. Clin. Microbiol."},{"key":"B42","doi-asserted-by":"publisher","first-page":"e00043","DOI":"10.1128\/mSystems.00043-17","article-title":"The Skin Microbiome of Cohabiting Couples","volume":"2","author":"Ross","year":"2017","journal-title":"mSystems"},{"key":"B43","doi-asserted-by":"publisher","first-page":"R60","DOI":"10.1186\/gb-2011-12-6-r60","article-title":"Metagenomic Biomarker Discovery and Explanation","volume":"12","author":"Segata","year":"2011","journal-title":"Gen. Biol."},{"key":"B44","first-page":"806","article-title":"Relationship of Abnormal Vaginal Flora, Proinflammatory Cytokines and Idiopathic Infertility in Women Undergoing IVF","volume":"46","author":"Spandorfer","year":"2001","journal-title":"J. Reprod. Med."},{"key":"B45","first-page":"123","article-title":"The Role of Lactotransferrin in the Molecular Mechanisms of Antimicrobial Defense","volume":"19","author":"Spik","year":"1983","journal-title":"Bull. Eur. Physiopathol. Respir."},{"key":"B46","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1186\/s12958-019-0562-z","article-title":"The Complex Microbiome From Native Semen to Embryo Culture Environment in Human In Vitro Fertilization Procedure","volume":"18","author":"\u0160t\u0161epetova","year":"2020","journal-title":"Reprod. Biol. Endocrinol."},{"key":"B47","doi-asserted-by":"publisher","first-page":"990","DOI":"10.1016\/S1701-2163(15)30412-6","article-title":"The Effect of Age, Ethnicity, and Level of Education on Fertility Awareness and Duration of Infertility","volume":"36","author":"Swift","year":"2014","journal-title":"J. Obstet Gynaecol Can."},{"key":"B48","doi-asserted-by":"publisher","first-page":"405","DOI":"10.1136\/sextrans-2017-053376","article-title":"Risk Factors for Mycoplasma Genitalium Endometritis and Incident Infection: A Secondary Data Analysis of the T-Cell Response Against Chlamydia (TRAC) Study","volume":"2","author":"Taylor","year":"2018","journal-title":"Sex Trans. Dis."},{"key":"B49","doi-asserted-by":"publisher","first-page":"883","DOI":"10.1016\/j.rbmo.2018.12.019","article-title":"Vaginal Microbiota Profile at the Time of Embryo Transfer Does Not Affect Live Birth Rate in IVF Cycles With Donated Oocytes","volume":"38","author":"Vergaro","year":"2019","journal-title":"Reprod. BioMed. Online"},{"key":"B50","doi-asserted-by":"publisher","first-page":"674","DOI":"10.1016\/S1472-6483(10)61683-5","article-title":"Vaginal Lactobacilli, Probiotics and IVF","volume":"11","author":"Verstraelen","year":"2005","journal-title":"Reprod. BioMed. Online"},{"key":"B51","doi-asserted-by":"publisher","first-page":"e110152","DOI":"10.1371\/journal.pone.0110152","article-title":"Bacterial Communities in Semen From Men of Infertile Couples: Metagenomic Sequencing Reveals Relationships of Seminal Microbiota to Semen Quality","volume":"9","author":"Weng","year":"2014","journal-title":"PloS One"},{"key":"B52","doi-asserted-by":"publisher","first-page":"714","DOI":"10.1111\/j.1471-0528.2002.01297.x","article-title":"Rates of Bacterial Vaginosis in Women Undergoing In Vitro Fertilisation for Different Types of Infertility","volume":"109","author":"Wilson","year":"2002","journal-title":"BJOG"},{"key":"B53","doi-asserted-by":"publisher","first-page":"135e9","DOI":"10.1016\/j.gyobfe.2003.11.017","article-title":"Abnormal Bacterial Colonisation of the Vagina and Implantation During Assisted Reproduction","volume":"32","author":"Wittemer","year":"2004","journal-title":"Gynecol. Obstet. Fertil."},{"key":"B54","doi-asserted-by":"publisher","first-page":"1613","DOI":"10.1099\/ijsem.0.001755","article-title":"Introducing EzBioCloud: A Taxonomically United Database of 16S rRNA and Whole Genome Assemblies","volume":"67","author":"Yoon","year":"2017","journal-title":"Int. J. Syst. Evol. Microbiol."}],"container-title":["Frontiers in Cellular and Infection Microbiology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2021.709372\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,10,1]],"date-time":"2021-10-01T12:23:38Z","timestamp":1633091018000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcimb.2021.709372\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,10,1]]},"references-count":54,"alternative-id":["10.3389\/fcimb.2021.709372"],"URL":"https:\/\/doi.org\/10.3389\/fcimb.2021.709372","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2020.10.22.20215988","asserted-by":"object"}]},"ISSN":["2235-2988"],"issn-type":[{"value":"2235-2988","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,10,1]]},"article-number":"709372"}}