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Is amniotic fluid of women with uncomplicated term pregnancies free of bacteria?

      Background

      The “sterile womb” paradigm is debated. Recent evidence suggests that the offspring’s first microbial encounter is before birth in term uncomplicated pregnancies. The establishment of a healthy microbiota early in life might be crucial for reducing the burden of diseases later in life.

      Objective

      We aimed to investigate the presence of a microbiota in sterilely collected amniotic fluid in uncomplicated pregnancies at term in the Preventing Atopic Dermatitis and Allergies in children (PreventADALL) study cohort.

      Study Design

      Amniotic fluid was randomly sampled at cesarean deliveries in pregnant women in 1 out of 3 study sites included in the PreventADALL study. From 65 pregnancies at term, where amniotic fluid was successfully sampled, we selected 10 from elective (planned, without ongoing labor) cesarean deliveries with intact amniotic membranes and all 14 with prior rupture of membranes were included as positive controls. Amniotic fluid was analyzed by culture-independent and culture-dependent techniques.

      Results

      The median (min-max) concentration of prokaryotic DNA (16S rRNA gene copies/mL; digital droplet polymerase chain reaction) was low for the group with intact membranes [664 (544–748)]–corresponding to the negative controls [596 (461–679)], while the rupture of amniotic membranes group had >10-fold higher levels [7700 (1066–251,430)] (P = .0001, by Mann-Whitney U test). Furthermore, bacteria were detected in 50% of the rupture of amniotic membranes samples by anaerobic culturing, while none of the intact membranes samples showed bacterial growth. Sanger sequencing of the rupture of amniotic membrane samples identified bacterial strains that are commonly part of the vaginal flora and/or associated with intrauterine infections.

      Conclusion

      We conclude that fetal development in uncomplicated pregnancies occurs in the absence of an amniotic fluid microbiota and that the offspring microbial colonization starts after uterine contractions and rupture of amniotic membrane.

      Key words

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      References

        • Lloyd-Price J.
        • Abu-Ali G.
        • Huttenhower C.
        The healthy human microbiome.
        Genome Med. 2016; 8: 51
        • Human Microbiome Project Consortium
        Structure, function and diversity of the healthy human microbiome.
        Nature. 2012; 486: 207-214
        • Amenyogbe N.
        • Kollmann T.R.
        • Ben-Othman R.
        Early-life host-microbiome interphase: the key frontier for immune development.
        Front Pediatr. 2017; 5: 111
        • Jenmalm M.C.
        The mother-offspring dyad: microbial transmission, immune interactions and allergy development.
        J Intern Med. 2017; 282: 484-495
        • Charbonneau M.R.
        • Blanton L.V.
        • DiGiulio D.B.
        • et al.
        A microbial perspective of human developmental biology.
        Nature. 2016; 535: 48-55
        • Stiemsma L.T.
        • Michels K.B.
        The role of the microbiome in the developmental origins of health and disease.
        Pediatrics. 2018; 141
        • Collado M.C.
        • Rautava S.
        • Aakko J.
        • Isolauri E.
        • Salminen S.
        Human gut colonization may be initiated in utero by distinct microbial communities in the placenta and amniotic fluid.
        Sci Rep. 2016; 6: 23129
        • Stroup P.E.
        Amniotic fluid infection and the intact fetal membrane.
        Obstet Gynecol. 1962; 19: 736-739
        • Lewis J.F.
        • Johnson P.
        • Miller P.
        Evaluation of amniotic fluid for aerobic and anaerobic bacteria.
        Am J Clin Pathol. 1976; 65: 58-63
        • Miller Jr., J.M.
        • Pupkin M.J.
        • Hill G.B.
        Bacterial colonization of amniotic fluid from intact fetal membranes.
        Am J Obstet Gynecol. 1980; 136: 796-804
        • Aagaard K.
        • Ma J.
        • Antony K.M.
        • Ganu R.
        • Petrosino J.
        • Versalovic J.
        The placenta harbors a unique microbiome.
        Sci Transl Med. 2014; 6: 237ra65
        • Doyle R.M.
        • Harris K.
        • Kamiza S.
        • et al.
        Bacterial communities found in placental tissues are associated with severe chorioamnionitis and adverse birth outcomes.
        PLoS One. 2017; 12: e0180167
        • Perez-Munoz M.E.
        • Arrieta M.C.
        • Ramer-Tait A.E.
        • Walter J.
        A critical assessment of the “sterile womb” and “in utero colonization” hypotheses: implications for research on the pioneer infant microbiome.
        Microbiome. 2017; 5: 48
        • Hornef M.
        • Penders J.
        Does a prenatal bacterial microbiota exist?.
        Mucosal Immunol. 2017; 10: 598-601
        • Lauder A.P.
        • Roche A.M.
        • Sherrill-Mix S.
        • et al.
        Comparison of placenta samples with contamination controls does not provide evidence for a distinct placenta microbiota.
        Microbiome. 2016; 4: 29
        • Carlsen K.C.L.
        • Rehbinder E.M.
        • Skjerven H.O.
        • et al.
        Preventing atopic dermatitis and allergies in children–the PreventADALL study.
        Allergy. 2018 Apr 30; (https://doi.org/10.1111/all.13468. [Epub ahead of print])
        • Hindson C.M.
        • Chevillet J.R.
        • Briggs H.A.
        • et al.
        Absolute quantification by droplet digital PCR versus analog real-time PCR.
        Nat Methods. 2013; 10: 1003-1005
        • Yu Y.
        • Lee C.
        • Kim J.
        • Hwang S.
        Group-specific primer and probe sets to detect methanogenic communities using quantitative real-time polymerase chain reaction.
        Biotechnol Bioeng. 2005; 89: 670-679
        • Caporaso J.G.
        • Kuczynski J.
        • Stombaugh J.
        • et al.
        QIIME allows analysis of high-throughput community sequencing data.
        Nat Methods. 2010; 7: 335-336
        • Edgar R.C.
        Search and clustering orders of magnitude faster than BLAST.
        Bioinformatics. 2010; 26: 2460-2461
        • Edgar R.C.
        UPARSE: highly accurate OTU sequences from microbial amplicon reads.
        Nat Methods. 2013; 10: 996-998
        • Quast C.
        • Pruesse E.
        • Yilmaz P.
        • et al.
        The SILVA ribosomal RNA gene database project: improved data processing and web-based tools.
        Nucleic Acids Res. 2013; 41: D590-D596
        • Salter S.J.
        • Cox M.J.
        • Turek E.M.
        • et al.
        Reagent and laboratory contamination can critically impact sequence-based microbiome analyses.
        BMC Biol. 2014; 12: 87
        • Seong H.S.
        • Lee S.E.
        • Kang J.H.
        • Romero R.
        • Yoon B.H.
        The frequency of microbial invasion of the amniotic cavity and histologic chorioamnionitis in women at term with intact membranes in the presence or absence of labor.
        Am J Obstet Gynecol. 2008; 199: 375.e1-375.e5
        • Kim M.J.
        • Romero R.
        • Gervasi M.T.
        • et al.
        Widespread microbial invasion of the chorioamniotic membranes is a consequence and not a cause of intra-amniotic infection.
        Lab Invest. 2009; 89: 924-936
        • Dominguez-Bello M.G.
        • Costello E.K.
        • Contreras M.
        • et al.
        Delivery mode shapes the acquisition and structure of the initial microbiota across multiple body habitats in newborns.
        Proc Natl Acad Sci U S A. 2010; 107: 11971-11975
        • Backhed F.
        • Roswall J.
        • Peng Y.
        • et al.
        Dynamics and stabilization of the human gut microbiome during the first year of life.
        Cell Host Microbe. 2015; 17: 852
        • Chu D.M.
        • Ma J.
        • Prince A.L.
        • Antony K.M.
        • Seferovic M.D.
        • Aagaard K.M.
        Maturation of the infant microbiome community structure and function across multiple body sites and in relation to mode of delivery.
        Nat Med. 2017; 23: 314-326
        • DiGiulio D.B.
        Diversity of microbes in amniotic fluid.
        Semin Fetal Neonatal Med. 2012; 17: 2-11
        • Shin H.
        • Pei Z.
        • Martinez II, K.A.
        • et al.
        The first microbial environment of infants born by C-section: the operating room microbes.
        Microbiome. 2015; 3: 59
        • Ravel J.
        • Gajer P.
        • Abdo Z.
        • et al.
        Vaginal microbiome of reproductive-age women.
        Proc Natl Acad Sci U S A. 2011; 108: 4680-4687
        • Romero R.
        • Mazor M.
        • Morrotti R.
        • et al.
        Infection and labor. VII. Microbial invasion of the amniotic cavity in spontaneous rupture of membranes at term.
        Am J Obstet Gynecol. 1992; 166: 129-133
        • Lannon S.M.R.
        • Adams Waldorf K.M.
        • Fiedler T.
        • et al.
        Parallel detection of lactobacillus and bacterial vaginosis-associated bacterial DNA in the chorioamnion and vagina of pregnant women at term.
        J Matern Fetal Neonatal Med. 2018; : 1-9
        • Lee S.M.
        • Lee K.A.
        • Kim S.M.
        • Park C.W.
        • Yoon B.H.
        The risk of intra-amniotic infection, inflammation and histologic chorioamnionitis in term pregnant women with intact membranes and labor.
        Placenta. 2011; 32: 516-521
        • Goldenberg R.L.
        • Culhane J.F.
        • Iams J.D.
        • Romero R.
        Epidemiology and causes of preterm birth.
        Lancet. 2008; 371: 75-84
        • Han Y.W.
        • Shen T.
        • Chung P.
        • Buhimschi I.A.
        • Buhimschi C.S.
        Uncultivated bacteria as etiologic agents of intra-amniotic inflammation leading to preterm birth.
        J Clin Microbiol. 2009; 47: 38-47
        • Combs C.A.
        • Gravett M.
        • Garite T.J.
        • et al.
        Amniotic fluid infection, inflammation, and colonization in preterm labor with intact membranes.
        Am J Obstet Gynecol. 2014; 210: 125.e1-125.e15
        • Dominguez-Bello M.G.
        • De Jesus-Laboy K.M.
        • Shen N.
        • et al.
        Partial restoration of the microbiota of cesarean-born infants via vaginal microbial transfer.
        Nat Med. 2016; 22: 250-253
        • Rowlands S.
        • Danielewski J.A.
        • Tabrizi S.N.
        • Walker S.P.
        • Garland S.M.
        Microbial invasion of the amniotic cavity in midtrimester pregnancies using molecular microbiology.
        Am J Obstet Gynecol. 2017; 217: 71.e1-71.e5
        • Pelzer E.
        • Gomez-Arango L.F.
        • Barrett H.L.
        • Nitert M.D.
        Review: Maternal health and the placental microbiome.
        Placenta. 2017; 54: 30-37
        • Parnell L.A.
        • Briggs C.M.
        • Mysorekar I.U.
        Maternal microbiomes in preterm birth: recent progress and analytical pipelines.
        Semin Perinatol. 2017; 41: 392-400
        • Prince A.L.
        • Ma J.
        • Kannan P.S.
        • et al.
        The placental membrane microbiome is altered among subjects with spontaneous preterm birth with and without chorioamnionitis.
        Am J Obstet Gynecol. 2016; 214: 627.e1-627.e16
        • Thornburg K.L.
        • Marshall N.
        The placenta is the center of the chronic disease universe.
        Am J Obstet Gynecol. 2015; 213: S14-S20