Microbial Ecology

, Volume 69, Issue 2, pp 407–414 | Cite as

Homogeneity of the Vaginal Microbiome at the Cervix, Posterior Fornix, and Vaginal Canal in Pregnant Chinese Women

  • Yi-E Huang
  • Yan Wang
  • Yan He
  • Yong Ji
  • Li-Ping Wang
  • Hua-Fang Sheng
  • Min Zhang
  • Qi-Tao Huang
  • Dong-Jing Zhang
  • Jing-Jing Wu
  • Mei ZhongEmail author
  • Hong-Wei ZhouEmail author


The vaginal microbiome is an emerging concern in prenatal health. Because the sampling process of vaginal microbiota may pose potential risks for pregnant women, the choice of sampling site should be carefully considered. However, whether the microbial diversity is different across various sampling sites has been controversial. In the present study, three repeated swabs were collected at the cervix (C), posterior fornix (P), and vaginal canal (V) from 34 Chinese women during different pregnancy stages, and vaginal species were determined using the Illumina sequencing of 16S rRNA tag sequences. The identified microbiomes were classified into four community state types (CSTs): CST I (dominated by L. crispatus), CST II (dominated by L. gasseri), CST III (dominated by L. iners), and CST IV-A (characterized by a low abundance of Lactobacillus, but with proportions of various species previously shown to be associated with bacterial vaginosis). All individuals had consistent CST at the three sampling sites regardless of pregnancy stage and CST group. In addition, there was little heterogeneity across community structures within each individual, as determined by LEfSe, indicating high vaginal microbiome homogeneity at the three sampling sites. The present study also revealed different beta diversity during pregnancy stages. The vaginal microbiome variation among women during trimester T1 (9 ± 2.6 weeks) is larger than that of non-pregnant women and women from other trimesters, as demonstrated by the UniFrac distance (P < 0.05). In particular, the present study is the first one that demonstrates the notably difference of vaginal microbiome of postpartum women compare to women in gestation. These results will be useful for future studies of the vaginal microbiota during pregnancy.


Lactobacillus Bacterial Vaginosis Terminal Restriction Fragment Length Polymorphism Vaginal Microbiota Vaginal Canal 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This work was supported through funding from the National Natural Science Foundation of China (NSFC31322003, 31270152), the Program for New Century Excellent Talents in University (NCET-11-0921), the Educational Commission of Guangdong Province of China (2012KJCX0031), and a grant from the School of Public Health and Tropical Medicine of Southern Medical University, China (Grant No. GW201316).

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  1. 1.
    Verstraelen H, Verhelst R, Claeys G, De Backer E, Temmerman M, Vaneechoutte M (2009) Longitudinal analysis of the vaginal microflora in pregnancy suggests that L. crispatus promotes the stability of the normal vaginal microflora and that L. gasseri and/or L. iners are more conducive to the occurrence of abnormal vaginal microflora. BMC Microbiol 116Google Scholar
  2. 2.
    Hernandez-Rodriguez C, Romero-Gonzalez R, Albani-Campanario M, Figueroa-Damian R, Meraz-Cruz N, Hernandez-Guerrero C (2011) Vaginal microbiota of healthy pregnant Mexican women is constituted by four Lactobacillus species and several vaginosis-associated bacteria. Infect Dis Obstet Gynecol 2011:851485PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Verhelst R, Verstraelen H, Claeys G, Verschraegen G, Van Simaey L, De Ganck C, De Backer E, Temmerman M, Vaneechoutte M (2005) Comparison between Gram stain and culture for the characterization of vaginal microflora: definition of a distinct grade that resembles grade I microflora and revised categorization of grade I microflora. BMC Microbiol 5:61PubMedCentralPubMedCrossRefGoogle Scholar
  4. 4.
    Hyman RW, Herndon CN, Jiang H, Palm C, Fukushima M, Bernstein D, Vo KC, Zelenko Z, Davis RW, Giudice LC (2012) The dynamics of the vaginal microbiome during infertility therapy with in vitro fertilization-embryo transfer. J Assist Reprod Genet 29:105–115PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Boskey ER, Cone RA, Whaley KJ, Moench TR (2001) Origins of vaginal acidity: high D/L lactate ratio is consistent with bacteria being the primary source. Hum Reprod 16:1809–1813PubMedCrossRefGoogle Scholar
  6. 6.
    Donders GG, Van Calsteren K, Bellen G, Reybrouck R, Van den Bosch T, Riphagen I, Van Lierde S (2009) Predictive value for preterm birth of abnormal vaginal flora, bacterial vaginosis and aerobic vaginitis during the first trimester of pregnancy. BJOG 116:1315–1324PubMedCrossRefGoogle Scholar
  7. 7.
    Hillier SL, Nugent RP, Eschenbach DA, Krohn MA, Gibbs RS, Martin DH, Cotch MF, Edelman R, Pastorek JG 2nd, Rao AV et al (1995) Association between bacterial vaginosis and preterm delivery of a low-birth-weight infant. The vaginal infections and prematurity study group. N Engl J Med 333:1737–1742PubMedCrossRefGoogle Scholar
  8. 8.
    Leitich H, Bodner-Adler B, Brunbauer M, Kaider A, Egarter C, Husslein P (2003) Bacterial vaginosis as a risk factor for preterm delivery: a meta-analysis. Am J Obstet Gynecol 189:139–147PubMedCrossRefGoogle Scholar
  9. 9.
    Gajer P, Brotman RM, Bai G, Sakamoto J, Schutte UM, Zhong X, Koenig SS, Fu L, Ma ZS, Zhou X, Abdo Z, Forney LJ, Ravel J (2012) Temporal dynamics of the human vaginal microbiota. Sci Transl Med 4:132ra152Google Scholar
  10. 10.
    Aagaard K, Riehle K, Ma J, Segata N, Mistretta TA, Coarfa C, Raza S, Rosenbaum S, Van den Veyver I, Milosavljevic A, Gevers D, Huttenhower C, Petrosino J, Versalovic J (2012) A metagenomic approach to characterization of the vaginal microbiome signature in pregnancy. PLoS ONE 7:e36466PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Romero R, Hassan SS, Gajer P, Tarca AL, Fadrosh DW, Nikita L, Galuppi M, Lamont RF, Chaemsaithong P, Miranda J, Chaiworapongsa T, Ravel J (2014) The composition and stability of the vaginal microbiota of normal pregnant women is different from that of non-pregnant women. Microbiome 2:4PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Cole AM (2006) Innate host defense of human vaginal and cervical mucosae. Curr Top Microbiol Immunol 306:199–230PubMedGoogle Scholar
  13. 13.
    Bartlett JG, Moon NE, Goldstein PR, Goren B, Onderdonk AB, Polk BF (1978) Cervical and vaginal bacterial flora: ecologic niches in the female lower genital tract. Am J Obstet Gynecol 130:658–661PubMedGoogle Scholar
  14. 14.
    Bartlett JG, Polk BF (1984) Bacterial flora of the vagina: quantitative study. Rev Infect Dis 6(Suppl 1):S67–S72PubMedCrossRefGoogle Scholar
  15. 15.
    Ling Z, Liu X, Chen X, Zhu H, Nelson KE, Xia Y, Li L, Xiang C (2011) Diversity of cervico vaginal microbiota associated with female lower genital tract infections. Microb Ecol 61:704–714PubMedCrossRefGoogle Scholar
  16. 16.
    Kim TK, Thomas SM, Ho M, Sharma S, Reich CI, Frank JA, Yeater KM, Biggs DR, Nakamura N, Stumpf R, Leigh SR, Tapping RI, Blanke SR, Slauch JM, Gaskins HR, Weisbaum JS, Olsen GJ, Hoyer LL, Wilson BA (2009) Heterogeneity of vaginal microbial communities within individuals. J Clin Microbiol 47:1181–1189PubMedCentralPubMedCrossRefGoogle Scholar
  17. 17.
    Forney LJ, Gajer P, Williams CJ, Schneider GM, Koenig SS, McCulle SL, Karlebach S, Brotman RM, Davis CC, Ault K, Ravel J (2010) Comparison of self-collected and physician-collected vaginal swabs for microbiome analysis. J Clin Microbiol 48:1741–1748PubMedCentralPubMedCrossRefGoogle Scholar
  18. 18.
    Zhou HW, Li DF, Tam NF, Jiang XT, Zhang H, Sheng HF, Qin J, Liu X, Zou F (2011) BIPES, a cost-effective high-throughput method for assessing microbial diversity. ISME J 5:741–749PubMedCentralPubMedCrossRefGoogle Scholar
  19. 19.
    Peng X, Yu KQ, Deng GH, Jiang YX, Wang Y, Zhang GX et al. (2013) Comparison of direct boiling method with commercial kits for extracting fecal microbiome DNA by Illumina sequencing of 16S rRNA tags. J Microbiol MethodsGoogle Scholar
  20. 20.
    Edgar RC, Haas BJ, Clemente JC, Quince C, Knight R (2011) UCHIME improves sensitivity and speed of chimera detection. Bioinformatics 27:2194–2200PubMedCentralPubMedCrossRefGoogle Scholar
  21. 21.
    Wang Q, Garrity GM, Tiedje JM, Cole JR (2007) Naive Bayesian classifier for rapid assignment of rRNA sequences into the new bacterial taxonomy. Appl Environ Microbiol 73:5261–5267PubMedCentralPubMedCrossRefGoogle Scholar
  22. 22.
    Lozupone C, Lladser ME, Knights D, Stombaugh J, Knight R (2011) UniFrac: an effective distance metric for microbial community comparison. ISME J 5:169–172PubMedCentralPubMedCrossRefGoogle Scholar
  23. 23.
    LozuponeC KR (2005) UniFrac: a new phylogenetic method for comparing microbial communities. Appl Environ Microbiol 71:8228–8235CrossRefGoogle Scholar
  24. 24.
    SPSS for Windows, Rel. 17.0.1. (2009). Chicago: SPSS Inc.Google Scholar
  25. 25.
    Segata N, Izard J, Waldron L, Gevers D, Miropolsky L, Garrett WS, Huttenhower C (2011) Metagenomic biomarker discovery and explanation. Genome Biol 12:R60PubMedCentralPubMedCrossRefGoogle Scholar
  26. 26.
    Zhou X, Brown CJ, Abdo Z, Davis CC, Hansmann MA, Joyce P, Foster JA, Forney LJ (2007) Differences in the composition of vaginal microbial communities found in healthy Caucasian and black women. ISME J 1:121–133PubMedCrossRefGoogle Scholar
  27. 27.
    Kiss H, Kogler B, Petricevic L, Sauerzapf I, Klayraung S, Domig K, Viernstein H, Kneifel W (2007) Vaginal Lactobacillus microbiota of healthy women in the late first trimester of pregnancy. BJOG 114:1402–1407PubMedCrossRefGoogle Scholar
  28. 28.
    Bezirtzoglou E, Voidarou C, Papadaki A, Tsiotsias A, Kotsovolou O, Konstandi M (2008) Hormone therapy alters the composition of the vaginal microflora in ovariectomized rats. Microb Ecol 55:751–759PubMedCrossRefGoogle Scholar
  29. 29.
    Ngai FW, Ngu SF (2014) Predictors of family and marital functioning at early postpartum. J Adv NursGoogle Scholar
  30. 30.
    Kalra A, Palcu CT, Sobel JD, Akins RA (2007) Bacterial vaginosis: culture- and PCR-based characterizations of a complex polymicrobial disease’s pathobiology. Curr Infect Dis Rep 9:485–500PubMedCrossRefGoogle Scholar
  31. 31.
    Ferris MJ, Norori J, Zozaya-Hinchliffe M, Martin DH (2007) Cultivation-independent analysis of changes in bacterial vaginosis flora following metronidazole treatment. J Clin Microbiol 45:1016–1018PubMedCentralPubMedCrossRefGoogle Scholar
  32. 32.
    Jakobsson T, Forsum U (2007) Lactobacillus iners: a marker of changes in the vaginal flora? J Clin Microbiol 45:3145PubMedCentralPubMedCrossRefGoogle Scholar
  33. 33.
    Fashemi B, Delaney ML, Onderdonk AB, Fichorova RN (2013) Effects of feminine hygiene products on the vaginal mucosal biome. Microbiol Ecol Health Dis 24Google Scholar
  34. 34.
    Gupta S, Kumar N, Singhal N, Kaur R, Manektala U (2006) Vaginal microflora in postmenopausal women on hormone replacement therapy. Indian J Pathol Microbiol 49:457–461PubMedGoogle Scholar
  35. 35.
    Gondo F, da Silva MG, Polettini J, Tristao Ada R, Peracoli JC, Witkin SS, Rudge MV (2011) Vaginal flora alterations and clinical symptoms in low-risk pregnant women. Gynecol Obstet Investig 71:158–162CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Yi-E Huang
    • 1
    • 3
  • Yan Wang
    • 2
  • Yan He
    • 1
  • Yong Ji
    • 1
  • Li-Ping Wang
    • 2
  • Hua-Fang Sheng
    • 1
  • Min Zhang
    • 2
  • Qi-Tao Huang
    • 2
  • Dong-Jing Zhang
    • 1
  • Jing-Jing Wu
    • 1
  • Mei Zhong
    • 2
    Email author
  • Hong-Wei Zhou
    • 1
    Email author
  1. 1.State Key Laboratory of Organ Failure Research, Department of Environmental Health, School of Public Health and Tropical MedicineSouthern Medical UniversityGuangzhouChina
  2. 2.Department of Obstetrics and Gynecology, NanFang HospitalSouthern Medical UniversityGuangzhouChina
  3. 3.Hunan University of MedicineHuaihuaChina

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