Analysis of Vaginal Lactic Acid Producing Bacteria in Healthy Women

  • Nam, Hye-Ran (Culture Collection of Antimicrobial Resistant Microbes, Department of Biology, Seoul Women's University) ;
  • Whang, Kyung-Hee (Dr. Whang's Clinic) ;
  • Lee, Yeon-Hee (Culture Collection of Antimicrobial Resistant Microbes, Department of Biology, Seoul Women's University)
  • Published : 2007.12.31

Abstract

Vaginal lactic acid-producing bacteria of 80 pre-menopausal women were studied by isolation on Blood and DeMan-Rogosa-Sharpe agar, PCR with group-specific primers for Lactobacillus-denaturing gradient gel electrophoresis (DGGE), and PCR with specific primers for V3 region in 16S rRNA-temporal temperature gel electrophoresis (TTGE). Conventional isolation method on media detected only one lactobacillus (Lactobacillus brevis) while TTGE detected only Lactobacillus sp. DGGE detected seven Lactobacillus species; L. coleohominis, L. crispatus, L. iners, L. reuteri, L. rhamnosus, L. vaginalis, and Leuconostoc lactis. L. acidophilus and L. gasseri, which are prevalent in Western women, were not detected in Korean women. Furthermore, L. rhamnosus, Leuc. lactis, L. coleohominis, and Weissella cibaria, which were not previously reported in the vaginal microbiota of Korean women, were detected. The five most prevalent LABs in vaginal microbiota in Korean women were L. iners, Enterococcus faecalis, L. crispatus, Leuc. lactis, and W. cibaria.

Keywords

References

  1. Boskey, E.R., K.M. Telsch, K.J. Whaley, T.R. Moench, and R.A. Cone. 1999. Acid production by vaginal flora in vitro is consistent with the rate and extent of vaginal acidification. Infect. Immun. 67, 5170-5175
  2. Boyd, M.A., M.A. Antonio, and S.L. Hillier. 2005. Comparison of API 50 CH strips to whole-chromosomal DNA probes for identification of Lactobacillus species. J. Clin. Microbiol. 43, 5309-5311 https://doi.org/10.1128/JCM.43.10.5309-5311.2005
  3. Burton, J.P., E. Devillard, P.A. Cadieux, J.A. Hammond, and G. Reid. 2004. Detection of Atopobium vaginae in postmenopausal women by cultivation-independent methods warrants further investigation. J. Clin. Microbiol. 42, 1829-1831 https://doi.org/10.1128/JCM.42.4.1829-1831.2004
  4. Chang, C.E., S.I. Pavlova, L. Tao, E.L. Kim, S.C. Kim, H.S. Yun, and J.S. So. 2002. Molecular identification of vaginal Lactobacillus spp. isolated from Korean women. J. Microbiol. Biotechnol. 12, 312-317
  5. Choi, H.J., C.I. Cheigh, S.B. Kim, J.C. Lee, D.W. Lee, S.W. Choi, J.M. Park, and Y.R. Pyun. 2002. Weissella kimchii sp. nov., a novel lactic acid bacterium from kimchi. Int. J. Syst. Evol. Microbiol. 52, 507-511 https://doi.org/10.1099/00207713-52-2-507
  6. Coolen, M.J., E. Post, C.C. Davis, and L.J. Forney. 2005. Characterization of microbial communities found in the human vagina by analysis of terminal restriction fragment length polymorphisms of 16S rRNA genes. Appl. Environ. Microbiol. 71, 8729-8737 https://doi.org/10.1128/AEM.71.12.8729-8737.2005
  7. Ennahar, S. and Y. Cai. 2002. Genetic evidence that Weissella kimchii Choi et al. 2002 is a later heterotypic synonym of Weissella cibaria Bjorkroth et al. 2002. Int. J. Syst. Evol. Microbiol. 54, 463-465 https://doi.org/10.1099/ijs.0.02783-0
  8. Giorgi, A., S. Torriani, F. Dellaglio, G. Bo, E. Stola, and L. Bernuzzi. 1987. Identification of vaginal lactobacilli from asymptomatic women. Microbiologica 10, 377-384
  9. Hoesl, C.E. and J.E. Altwein. 2005. The probiotic approach: an alternative treatment option in urology. Eur. Urol. 47, 288-296 https://doi.org/10.1016/j.eururo.2004.09.011
  10. Jang, J., B. Kim, J. Lee, J. Kim, G. Jeong, and H. Han. 2002. Identification of Weissella species by the genus-specific amplified ribosomal DNA restriction analysis. FEMS Microbiol. Lett. 212, 29-34 https://doi.org/10.1111/j.1574-6968.2002.tb11240.x
  11. Lachlak, N., E. Ageron, O. Zampatti, G. Michel, and P.A. Grimont. 1996. Composition of the Lactobacillus acidophilus complex isolated from vaginal flora. New Microbiol. 19, 123-132
  12. Lane, D.J. 1991. 16S/23S rRNA sequencing, p. 115-175. In E.R. Stackebrandt and M. Goodfellow (ed.), Nucleic acid techniques in bacterial systematics. John Wiley and Sons Ltd., Chichester, UK
  13. Martin, H.L., B.A. Richardson, P.M. Nyange, L. Lavreys, S.L. Hillier, B. Chohan, K. Mandaliya, J.O. Ndinya-Achola, J. Bwayo, and J. Kreiss. 1999. Vaginal lactobacilli, microbial flora, and risk of human immunodeficiency virus type 1 and sexually transmitted disease acquisition. J. Infect. Dis. 180, 1863-1868 https://doi.org/10.1086/315127
  14. Muyzer, G., E.C. De Waal, and A.G. Uitterlinden. 1993. Profiling of complex microbial populations by denaturing gradient gel electrophoresis analysis of polymerase chain reaction-amplified genes coding for 16S rRNA. Appl. Environ. Microbiol. 59, 695-700
  15. Nikolaitchouk, N., C. Wacher, E. Falsen, B. Andersch, M.D. Collins, and P.A. Lawson. 2001. Lactobacillus coleohominis sp. nov., isolated from human sources. Int. J. Syst. Evol. Microbiol. 51, 2081-2085 https://doi.org/10.1099/00207713-51-6-2081
  16. Ogier, J.C., O. Son, A. Gruss, P. Tailliez, and A. Delacroix-Buchet. 2002. Identification of the bacterial microbiota in dairy products by temporal temperature gradient gel electrophoresis. Appl. Environ. Microbiol. 68, 3691-3701 https://doi.org/10.1128/AEM.68.8.3691-3701.2002
  17. Onderdonk, A.B. and K.W. Wissemann. 1993. Normal vaginal microbiota, p. 285-304. In P. Elsner and J. Martins (ed.), Vulvovaginitis. Marcel Dekker Inc., New York, N.Y., USA
  18. O'Sullivan, M.G., G.C. Thornton, G.C. O'Sullivan, and J.K. Collins. 1992. Probiotic bacteria myth or reality? Trends Food Sci. 31, 309-314
  19. Redondo-Lopez, V., R.L. Cook, and J.D. Sobel. 1990. Emerging role of lactobacilli in the control and maintenance of the vaginal bacterial microflora. Rev. Infect. Dis. 12, 856-872 https://doi.org/10.1093/clinids/12.5.856
  20. Reid, G., J.A. McGroarty, L. Tomeczek, and A.W. Bruce. 1996. Identification and plasmid profiles of Lactobacillus species from the vagina of 100 healthy women. FEMS Immunol. Med. Microbiol. 15, 23-26 https://doi.org/10.1111/j.1574-695X.1996.tb00354.x
  21. Sanger, F., S. Nicklen, and A.R. Coulson. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA 74, 5463-5467
  22. Silvester, M.E. and L.M. Dicks. 2003. Identification of lactic acid bacteria isolated from human vaginal secretions. Antonie Van Leeuwenhoek 83, 117-123 https://doi.org/10.1023/A:1023373023115
  23. Song, Y.L., N. Kato, Y. Matsumiya, C.X. Liu, H. Kato, and K. Watanabe. 1999. Identification of and hydrogen peroxide production by fecal and vaginal lactobacilli isolated from Japanese women and newborn infants. J. Clin. Microbiol. 37, 3062-3064
  24. Steyn, P.L. and W.H. Holzaphel. 1991. Identity and antibiotic sensitivity of human vaginal lactic acid bacteria from the population of northern Namibia. S-Afr. Tydskr. Wet. 87, 68-69
  25. Walter, J., C. Hertel, G.W. Tannock, C.M. Lis, K. Munro, and W.P. Hammes. 2001. Detection of Lactobacillus, Pediococcus, Leuconostoc, and Weissella species in human feces by using groupspecific PCR primers and denaturing gradient gel electrophoresis. Appl. Environ. Microbiol. 67, 2578-2585 https://doi.org/10.1128/AEM.67.6.2578-2585.2001
  26. Wilks, M., R. Wiggins, A. Whiley, E. Hennessy, S. Warwick, H. Porter, A. Corfield, and M. Millar. 2004. Identification and $H_2O_2$ production of vaginal Lactobacilli from pregnant women at high risk of preterm birth and relation with outcome. J. Clin. Microbiol. 42, 713-717 https://doi.org/10.1128/JCM.42.2.713-717.2004
  27. Zhou, X., S.J. Bent, M.G. Schneider, C.C. Davis, M.R. Islam, and L.J. Forney. 2004. Characterization of vaginal microbial communities in adult healthy women using cultivation-independent methods. Microbiology 150, 2565-2573 https://doi.org/10.1099/mic.0.26905-0