1887

Abstract

The present study aimed to investigate the prevalence and quantity of species in the vaginas of healthy Chinese women. Vaginal samples from 92 fertile and 22 postmenopausal healthy Chinese women were analysed using a 16S rRNA gene clone library and species-specific PCR followed by sequencing and real-time PCR. A total of 13 different species were detected. Species-specific PCR showed that 3 % of the fertile women were colonized by one species of and 97 % were colonized by two or more species. Among the postmenopausal women, 91 % were colonized by one species of and 9 % were colonized by two species. In the fertile women, (82.61 %), (70.65 %) and (67.39 %) were more prevalent than (40.22 %), (39.13 %), (23.91 %), (5.43 %), (3.26 %), (2.17 %), (2.17 %), (1.09 %), (1.09 %) and (1.09 %); was not detected. In the postmenopausal women, , , and were not detected, and the other 10 species were detected in just a few samples. The prevalence of these species according to the clone library differed from the prevalence indicated by the species-specific PCR. According to the semiquantitative analysis, the total DNA concentrations were higher in fertile women than in postmenopausal women. Sixty-one per cent of the fertile women were predominantly colonized by , 35 % by , and 2 % by . Associations between pairs of species in fertile women were significant (<0.05) between the following pairs: and , and , and , and , and and . In conclusion, this study provided detailed information on species colonizing the vaginas of healthy Chinese fertile and postmenopausal women. The study also showed that the diversity of species in fertile women was higher than in postmenopausal women. According to our study, different techniques, such as species-specific PCR and comparison against a 16S rRNA gene clone library, resulted in different findings regarding species prevalence. These findings highlight the importance of standardization of techniques used for evaluation of bacterial communities. According to our findings regarding species associations, and may have influences on colonization and proliferation of other vaginal species.

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.038687-0
2012-05-01
2019-12-06
Loading full text...

Full text loading...

/deliver/fulltext/jmm/61/5/729.html?itemId=/content/journal/jmm/10.1099/jmm.0.038687-0&mimeType=html&fmt=ahah

References

  1. Antonio M. A., Hawes S. E., Hillier S. L.. ( 1999;). The identification of vaginal Lactobacillus species and the demographic and microbiologic characteristics of women colonized by these species. . J Infect Dis 180:, 1950–1956. [CrossRef][PubMed]
    [Google Scholar]
  2. Antonio M. A., Meyn L. A., Murray P. J., Busse B., Hillier S. L.. ( 2009;). Vaginal colonization by probiotic Lactobacillus crispatus CTV-05 is decreased by sexual activity and endogenous lactobacilli. . J Infect Dis 199:, 1506–1513. [CrossRef][PubMed]
    [Google Scholar]
  3. Anukam K., Osazuwa E., Ahonkhai I., Ngwu M., Osemene G., Bruce A. W., Reid G.. ( 2006;). Augmentation of antimicrobial metronidazole therapy of bacterial vaginosis with oral probiotic Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14: randomized, double-blind, placebo controlled trial. . Microbes Infect 8:, 1450–1454. [CrossRef][PubMed]
    [Google Scholar]
  4. Bolton M., van der Straten A., Cohen C. R.. ( 2008;). Probiotics: potential to prevent HIV and sexually transmitted infections in women. . Sex Transm Dis 35:, 214–225. [CrossRef][PubMed]
    [Google Scholar]
  5. Boskey E. R., Telsch K. M., Whaley K. J., Moench T. R., Cone R. A.. ( 1999;). Acid production by vaginal flora in vitro is consistent with the rate and extent of vaginal acidification. . Infect Immun 67:, 5170–5175.[PubMed]
    [Google Scholar]
  6. Bradshaw C. S., Morton A. N., Hocking J., Garland S. M., Morris M. B., Moss L. M., Horvath L. B., Kuzevska I., Fairley C. K.. ( 2006a;). High recurrence rates of bacterial vaginosis over the course of 12 months after oral metronidazole therapy and factors associated with recurrence. . J Infect Dis 193:, 1478–1486. [CrossRef][PubMed]
    [Google Scholar]
  7. Bradshaw C. S., Tabrizi S. N., Fairley C. K., Morton A. N., Rudland E., Garland S. M.. ( 2006b;). The association of Atopobium vaginae and Gardnerella vaginalis with bacterial vaginosis and recurrence after oral metronidazole therapy. . J Infect Dis 194:, 828–836. [CrossRef][PubMed]
    [Google Scholar]
  8. Burton J. P., Reid G.. ( 2002;). Evaluation of the bacterial vaginal flora of 20 postmenopausal women by direct (Nugent score) and molecular (polymerase chain reaction and denaturing gradient gel electrophoresis) techniques. . J Infect Dis 186:, 1770–1780. [CrossRef][PubMed]
    [Google Scholar]
  9. Burton J. P., Cadieux P. A., Reid G.. ( 2003;). Improved understanding of the bacterial vaginal microbiota of women before and after probiotic instillation. . Appl Environ Microbiol 69:, 97–101. [CrossRef][PubMed]
    [Google Scholar]
  10. Byun R., Nadkarni M. A., Chhour K. L., Martin F. E., Jacques N. A., Hunter N.. ( 2004;). Quantitative analysis of diverse Lactobacillus species present in advanced dental caries. . J Clin Microbiol 42:, 3128–3136. [CrossRef][PubMed]
    [Google Scholar]
  11. Cadieux P. A., Burton J., Devillard E., Reid G.. ( 2009;). Lactobacillus by-products inhibit the growth and virulence of uropathogenic Escherichia coli. . J Physiol Pharmacol 60: (Suppl. 6), 13–18.[PubMed]
    [Google Scholar]
  12. Cauci S., Driussi S., De Santo D., Penacchioni P., Iannicelli T., Lanzafame P., De Seta F., Quadrifoglio F., de Aloysio D., Guaschino S.. ( 2002;). Prevalence of bacterial vaginosis and vaginal flora changes in peri- and postmenopausal women. . J Clin Microbiol 40:, 2147–2152. [CrossRef][PubMed]
    [Google Scholar]
  13. De Backer E., Verhelst R., Verstraelen H., Alqumber M. A., Burton J. P., Tagg J. R., Temmerman M., Vaneechoutte M.. ( 2007;). Quantitative determination by real-time PCR of four vaginal Lactobacillus species, Gardnerella vaginalis and Atopobium vaginae indicates an inverse relationship between L. gasseri and L. iners. . BMC Microbiol 7:, 115. [CrossRef][PubMed]
    [Google Scholar]
  14. Devillard E., Burton J. P., Hammond J. A., Lam D., Reid G.. ( 2004;). Novel insight into the vaginal microflora in postmenopausal women under hormone replacement therapy as analyzed by PCR-denaturing gradient gel electrophoresis. . Eur J Obstet Gynecol Reprod Biol 117:, 76–81. [CrossRef][PubMed]
    [Google Scholar]
  15. Dewhirst F. E., Chen T., Izard J., Paster B. J., Tanner A. C., Yu W. H., Lakshmanan A., Wade W. G.. ( 2010;). The human oral microbiome. . J Bacteriol 192:, 5002–5017. [CrossRef][PubMed]
    [Google Scholar]
  16. Ehrström S., Daroczy K., Rylander E., Samuelsson C., Johannesson U., Anzén B., Påhlson C.. ( 2010;). Lactic acid bacteria colonization and clinical outcome after probiotic supplementation in conventionally treated bacterial vaginosis and vulvovaginal candidiasis. . Microbes Infect 12:, 691–699. [CrossRef][PubMed]
    [Google Scholar]
  17. Falsen E., Pascual C., Sjödén B., Ohlén M., Collins M. D.. ( 1999;). Phenotypic and phylogenetic characterization of a novel Lactobacillus species from human sources: description of Lactobacillus iners sp. nov.. Int J Syst Bacteriol 49:, 217–221. [CrossRef][PubMed]
    [Google Scholar]
  18. Ferris M. J., Norori J., Zozaya-Hinchliffe M., Martin D. H.. ( 2007;). Cultivation-independent analysis of changes in bacterial vaginosis flora following metronidazole treatment. . J Clin Microbiol 45:, 1016–1018. [CrossRef][PubMed]
    [Google Scholar]
  19. Fredricks D. N., Fiedler T. L., Marrazzo J. M.. ( 2005;). Molecular identification of bacteria associated with bacterial vaginosis. . N Engl J Med 353:, 1899–1911. [CrossRef][PubMed]
    [Google Scholar]
  20. Garg K. B., Ganguli I., Das R., Talwar G. P.. ( 2009;). Spectrum of Lactobacillus species present in healthy vagina of Indian women. . Indian J Med Res 129:, 652–657.[PubMed]
    [Google Scholar]
  21. Heilig H. G., Zoetendal E. G., Vaughan E. E., Marteau P., Akkermans A. D., de Vos W. M.. ( 2002;). Molecular diversity of Lactobacillus spp. and other lactic acid bacteria in the human intestine as determined by specific amplification of 16S ribosomal DNA. . Appl Environ Microbiol 68:, 114–123. [CrossRef][PubMed]
    [Google Scholar]
  22. Heinemann C., Reid G.. ( 2005;). Vaginal microbial diversity among postmenopausal women with and without hormone replacement therapy. . Can J Microbiol 51:, 777–781. [CrossRef][PubMed]
    [Google Scholar]
  23. Hillier S. L., Lau R. J.. ( 1997;). Vaginal microflora in postmenopausal women who have not received estrogen replacement therapy. . Clin Infect Dis 25: (Suppl. 2), S123–S126. [CrossRef][PubMed]
    [Google Scholar]
  24. Hyman R. W., Fukushima M., Diamond L., Kumm J., Giudice L. C., Davis R. W.. ( 2005;). Microbes on the human vaginal epithelium. . Proc Natl Acad Sci U S A 102:, 7952–7957. [CrossRef][PubMed]
    [Google Scholar]
  25. Joesoef M., Schmid G.. ( 2003;). Bacterial vaginosis. . Clin Evid 10:, 1824–1833.[PubMed]
    [Google Scholar]
  26. Kemp P. F., Aller J. Y.. ( 2004;). Bacterial diversity in aquatic and other environments: what 16S rDNA libraries can tell us. . FEMS Microbiol Ecol 47:, 161–177. [CrossRef][PubMed]
    [Google Scholar]
  27. Kiss H., Kögler 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:, 1402–1407. [CrossRef][PubMed]
    [Google Scholar]
  28. Lamont R. F., Sobel J. D., Akins R. A., Hassan S. S., Chaiworapongsa T., Kusanovic J. P., Romero R.. ( 2011;). The vaginal microbiome: new information about genital tract flora using molecular based techniques. . BJOG 118:, 533–549. [CrossRef][PubMed]
    [Google Scholar]
  29. Ling Z., Kong J., Liu F., Zhu H., Chen X., Wang Y., Li L., Nelson K. E., Xia Y., Xiang C.. ( 2010;). Molecular analysis of the diversity of vaginal microbiota associated with bacterial vaginosis. . BMC Genomics 11:, 488. [CrossRef][PubMed]
    [Google Scholar]
  30. Linhares I. M., Giraldo P. C., Baracat E. C.. ( 2010;). [New findings about vaginal bacterial flora]. . Rev Assoc Med Bras 56:, 370–374 (in Portuguese). [CrossRef][PubMed]
    [Google Scholar]
  31. Liu J. M., Sun J., Yuan J. L., Kang B.. ( 2006;). Difference of lactobacillus flora of vagina between healthy women and women with bacteria vaginosis. . Prog Obstet Gynecol 115:, 18–20.
    [Google Scholar]
  32. Liu X. Q., Yang Y., Ma F., Zhang X. Y.. ( 2010;). Progress on the evaluation system of human vaginal micro-eco-environment. . Chin J Microecol 22:, 662–665.
    [Google Scholar]
  33. MacPhee R. A., Hummelen R., Bisanz J. E., Miller W. L., Reid G.. ( 2010;). Probiotic strategies for the treatment and prevention of bacterial vaginosis. . Expert Opin Pharmacother 11:, 2985–2995. [CrossRef][PubMed]
    [Google Scholar]
  34. Martinez R. C., Franceschini S. A., Patta M. C., Quintana S. M., Gomes B. C., De Martinis E. C., Reid G.. ( 2009;). Improved cure of bacterial vaginosis with single dose of tinidazole (2 g), Lactobacillus rhamnosus GR-1, and Lactobacillus reuteri RC-14: a randomized, double-blind, placebo-controlled trial. . Can J Microbiol 55:, 133–138. [CrossRef][PubMed]
    [Google Scholar]
  35. Pabich W. L., Fihn S. D., Stamm W. E., Scholes D., Boyko E. J., Gupta K.. ( 2003;). Prevalence and determinants of vaginal flora alterations in postmenopausal women. . J Infect Dis 188:, 1054–1058. [CrossRef][PubMed]
    [Google Scholar]
  36. Pavlova S. I., Kilic A. O., Kilic S. S., So J. S., Nader-Macias M. E., Simoes J. A., Tao L.. ( 2002;). Genetic diversity of vaginal lactobacilli from women in different countries based on 16S rRNA gene sequences. . J Appl Microbiol 92:, 451–459. [CrossRef][PubMed]
    [Google Scholar]
  37. Reid G.. ( 1999;). The scientific basis for probiotic strains of Lactobacillus. . Appl Environ Microbiol 65:, 3763–3766.[PubMed]
    [Google Scholar]
  38. Reid G.. ( 2006a;). Probiotics to prevent the need for, and augment the use of, antibiotics. . Can J Infect Dis Med Microbiol 17:, 291–295.[PubMed]
    [Google Scholar]
  39. Reid G.. ( 2006b;). Safe and efficacious probiotics: what are they?. Trends Microbiol 14:, 348–352. [CrossRef][PubMed]
    [Google Scholar]
  40. Reid G., Bruce A. W.. ( 2001;). Selection of lactobacillus strains for urogenital probiotic applications. . J Infect Dis 183: (Suppl. 1), S77–S80. [CrossRef][PubMed]
    [Google Scholar]
  41. Reid G., Bruce A. W., Fraser N., Heinemann C., Owen J., Henning B.. ( 2001;). Oral probiotics can resolve urogenital infections. . FEMS Immunol Med Microbiol 30:, 49–52. [CrossRef][PubMed]
    [Google Scholar]
  42. Reid G., Charbonneau D., Erb J., Kochanowski B., Beuerman D., Poehner R., Bruce A. W.. ( 2003;). Oral use of Lactobacillus rhamnosus GR-1 and L. fermentum RC-14 significantly alters vaginal flora: randomized, placebo-controlled trial in 64 healthy women. . FEMS Immunol Med Microbiol 35:, 131–134. [CrossRef][PubMed]
    [Google Scholar]
  43. Reid G., Younes J. A., Van der Mei H. C., Gloor G. B., Knight R., Busscher H. J.. ( 2011;). Microbiota restoration: natural and supplemented recovery of human microbial communities. . Nat Rev Microbiol 9:, 27–38. [CrossRef][PubMed]
    [Google Scholar]
  44. Robert H., Gabriel V., Fontagné-Faucher C.. ( 2009;). Biodiversity of lactic acid bacteria in French wheat sourdough as determined by molecular characterization using species-specific PCR. . Int J Food Microbiol 135:, 53–59. [CrossRef][PubMed]
    [Google Scholar]
  45. Shi Y., Chen L., Tong J., Xu C.. ( 2009;). Preliminary characterization of vaginal microbiota in healthy Chinese women using cultivation-independent methods. . J Obstet Gynaecol Res 35:, 525–532. [CrossRef][PubMed]
    [Google Scholar]
  46. Shiraishi T., Fukuda K., Morotomi N., Imamura Y., Mishima J., Imai S., Miyazawa K., Taniguchi H.. ( 2011;). Influence of menstruation on the microbiota of healthy women’s labia minora as analyzed using a 16S rRNA gene-based clone library method. . Jpn J Infect Dis 64:, 76–80.[PubMed]
    [Google Scholar]
  47. Sobel J. D., Schmitt C., Meriwether C.. ( 1993;). Long-term follow-up of patients with bacterial vaginosis treated with oral metronidazole and topical clindamycin. . J Infect Dis 167:, 783–784. [CrossRef][PubMed]
    [Google Scholar]
  48. Vallor A. C., Antonio M. A., Hawes S. E., Hillier S. L.. ( 2001;). Factors associated with acquisition of, or persistent colonization by, vaginal lactobacilli: role of hydrogen peroxide production. . J Infect Dis 184:, 1431–1436. [CrossRef][PubMed]
    [Google Scholar]
  49. Vásquez A., Jakobsson T., Ahrné S., Forsum U., Molin G.. ( 2002;). Vaginal Lactobacillus flora of healthy Swedish women. . J Clin Microbiol 40:, 2746–2749. [CrossRef][PubMed]
    [Google Scholar]
  50. Ventura M., Zink R.. ( 2002;). Specific identification and molecular typing analysis of Lactobacillus johnsonii by using PCR-based methods and pulsed-field gel electrophoresis. . FEMS Microbiol Lett 217:, 141–154. [CrossRef][PubMed]
    [Google Scholar]
  51. Vermeulen G. M., van Zwet A. A., Bruinse H. W.. ( 2001;). Changes in the vaginal flora after two percent clindamycin vaginal cream in women at high risk of spontaneous preterm birth. . BJOG 108:, 697–700. [CrossRef][PubMed]
    [Google Scholar]
  52. Walter J., Hertel C., Tannock G. W., Lis C. M., Munro K., Hammes W. P.. ( 2001;). Detection of Lactobacillus, Pediococcus, Leuconostoc, and Weissella species in human feces by using group-specific PCR primers and denaturing gradient gel electrophoresis. . Appl Environ Microbiol 67:, 2578–2585. [CrossRef][PubMed]
    [Google Scholar]
  53. Wang M., Song L., Yao L.H., Jia R., Chen H., Liu Y., Zhang Z.Q.. & Q, X.U. ( 2009;). Isolation and identification of vaginal lactobacilli from 223 healthy women of childbearing age. . Chin J Clin Obstet Gynecol 10:, 200–203.
    [Google Scholar]
  54. Zhou X., Bent S. J., Schneider M. G., Davis C. C., Islam M. R., Forney L. J.. ( 2004;). Characterization of vaginal microbial communities in adult healthy women using cultivation-independent methods. . Microbiology 150:, 2565–2573. [CrossRef][PubMed]
    [Google Scholar]
  55. Zozaya-Hinchliffe M., Lillis R., Martin D. H., Ferris M. J.. ( 2010;). Quantitative PCR assessments of bacterial species in women with and without bacterial vaginosis. . J Clin Microbiol 48:, 1812–1819. [CrossRef][PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.038687-0
Loading
/content/journal/jmm/10.1099/jmm.0.038687-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error