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Production of Antimicrobial Substances by Lactic Acid Bacteria I

Determination of Hydrogen Peroxide

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Public Health Microbiology

Abstract

Restoration of the balance of different ecological niches has been proposed as a way to control the income of pathogenic microorganisms (1). The genus Lactobacillus has been used in different human and animal tracts as probiotic microorganisms with this objective in mind (2). The characteristics of the strains proposed as probiotics have been published (3) or patented under the process of elaboration of different types of products (46). One of the mechanisms suggested to control the vaginal ecosystem is the production of antagonistic substances (lactic acid, bacteriocins, or H2O2) (79). The H2O2-producing microorganisms present in the vagina of healthy women have been suggested as some of the bacteria responsible for maintenance of ecological balance, mainly in pregnant women (10,11). The absence of these microorganisms is related to a higher risk of: bacterial vaginosis (12), recurrent urinary tract infections by Escherichia coli (13), and acquisition of human immunodeficiency virus type 1 (HIV-1) (14). Bauer (15) has proposed that H2O2-producing lactobacilli also might exert control over vaginal cancer through specific interactions of reactive oxygen species, such as superoxide anion, hydroxyl radicals, and hypochlorous acid. The conversion of H2O2 into more toxic compounds during the oxidative process is potentiated by peroxidase and halures. This enzyme and some halures, such as chloride and bromide, are present in vaginal washes in sufficient amounts (16) to allow an optimal environment for successful inhibition of pathogens.

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References

  1. Reid, G. (1999) The scientific basis for probiotic strains of Lactobacillus. App. Env. Microbiol. 65, 3763–3766.

    CAS  Google Scholar 

  2. Redondo-López, V., Cook, R. L., and Sobel, J. D. (1990) Emerging role of lactobacilli in the control and maintenance of the vaginal bacterial microflora. Rev. Infect. Dis. 12, 856–872.

    PubMed  Google Scholar 

  3. Reid, G. and Bruce, A. W. (2001) Selection of Lactobacillus strains for urogenital probiotic applications. J. Infect. Dis. 183(suppl 1), S77–S80.

    Article  PubMed  Google Scholar 

  4. Chrisope, G. L. (2000) Vaginal Lactobacillus medicant. US patent & trademark office, United States Patent, GyneLogix, Inc., Boulder, CO, No. 093,394-6-21.

    Google Scholar 

  5. Larsen, B. (1999) Vaginal pharmaceutical compositions. US patent & trademark office, United States Patent, Marshall University Research Corp. Huntington, WV, no. 5,958,461-7-34

    Google Scholar 

  6. Reid, G. and Bruce, A. W. (1997) Lactobacillus and skim milk compositions and methods for preventing microbial urogenital infections. US patent & trademark office, United States Patent, Research Corporation Technologies, Inc., Tucson, AZ, No. 5,643,830-1-1.

    Google Scholar 

  7. Eschenbach, D., Davick, P., Williams, S., Klebanoff, S., Young-Smith, C., and Holmes, K. K. (1989) Prevalence of hydrogen peroxide-producing Lactobacillus species in normal women with bacterial vaginosis. J. Clin. Microbiol. 27, 251–256.

    PubMed  CAS  Google Scholar 

  8. Jack, R. W., Tagg, J. R., and Ray, B. (1995) Bacteriocins of Gram-positive bacteria. Microbiol. Rev. 59, 171–200.

    PubMed  CAS  Google Scholar 

  9. Aroutcheva, A., Gariti, D., Simon, M., et al. (2001) Defense factors of vaginal lactobacilli. Am. J. Obstet. Gynecol. 185, 375–379.

    Article  PubMed  CAS  Google Scholar 

  10. McGroarty, J. (1993) Probiotic use of lactobacilli in the human female urogenital tract. FEMS Immunol. Med. Microbiol. 6, 251–264.

    Article  PubMed  CAS  Google Scholar 

  11. Goffeng, A. R., Holst, E., Milsom, I., Lindstedt, G., Lundberg, P. A., and Andersch, B. (1997) Fetal fibronectin and microorganisms in vaginal fluid of women with complicated pregnancies. Acta Obstet. Gynecol. Scand. 76, 521–527.

    Article  PubMed  CAS  Google Scholar 

  12. Hawes, S. E., Hillier, S. L., Benedetti, J., et al. (1996) Hydrogen peroxide-producing lactobacilli and acquisition of vaginal infections. J. Infec. Dis. 174, 1058–1063.

    CAS  Google Scholar 

  13. Gupta, K., Stapleton, A. E., Hooton, T. M., Roberts, P. L., Fennell, C. L., and Stamm, W. E. (1998) Inverse association of H2O2-producing lactobacilli and vaginal Escherichia coli colonization in women with recurrent urinary tract infections. J. Infect. Dis. 178, 446–450.

    PubMed  CAS  Google Scholar 

  14. Martin, H. L. Jr., Richardson, B. A., Nyange, P. M., et al. (1999) Vaginal lactobacilli, microbial flora, and risk of human immunodeficiency virus type 1 and sexually transmitted disease acquisition. J. Infect. Dis. 180, 1863–1868.

    Article  PubMed  CAS  Google Scholar 

  15. Bauer, G. (2001) Lactobacilli-mediated control of vaginal cancer through specific reactive oxygen species interaction. Med. Hypotheses. 57, 252–257.

    Article  PubMed  CAS  Google Scholar 

  16. Klebanoff, S. J., Hillier, S. L., Eschenbach, D. A., Waltersdorph, A. M. (1991) Control of the microbial flora of the vagina by H2O2-generating lactobacilli. J. Infect. Dis. 164, 94–100.

    PubMed  CAS  Google Scholar 

  17. Juven, B. J. and Pierson, M. D. (1996) Antibacterial effects of hydrogen peroxide and methods for its detection and quantification. J. Food Proteins 11, 1233–1241.

    Google Scholar 

  18. Berthier, F. (1993) On the screening of the hydrogen peroxide-generating lactic acid bacteria. Lett. Appl. Microbiol. 16, 150–153.

    Article  CAS  Google Scholar 

  19. Nuñez de Kairuz, M. S., Olazábal, M. E., Oliver, G., de Ruiz Holgado, A. P., and Farías, R. N. (1988) Fatty acid-dependent hydrogen peroxide production in Lactobacillus. Biochem. Biophys. Res. Commun. 152, 113–121.

    Article  PubMed  Google Scholar 

  20. Ocaña, V. S., Bru, E., de Ruiz Holgado, A. P., and Nader-Macías, M. E. (1999) Surface characteristics of lactobacilli isolated from human vagina. J. Gen. Appl. Microbiol. 45, 203–212.

    Article  PubMed  Google Scholar 

  21. Otero, C., Silva de Ruiz, C., Ibáñez, R., Wilde, O., Ruiz Holgado, A. and Nader-Macías, M. E. (1999) Lactobacilli and enterococci isolated from the bovine vaginal during the estrous cycle. Anaerobe 5, 305–307.

    Article  Google Scholar 

  22. Ocaña, V.S., P.de Ruiz Holgado, A., and Nader-Macías, M. E. (1999) Selection of vaginal H2O2-generating Lactobacillus for probiotic use. Curr. Microbiol. 38, 279–284.

    Article  PubMed  Google Scholar 

  23. Ocaña, V.S., P.de Ruiz Holgado, A., and Nader-Macías, M. E. (1999) Growth inhibition of Staphylococcus aureus by H2O2-producing Lactobacillus paracasei subsp. paracasei isolated from the human vagina. FEMS Immunol. Med. Microbiol. 23, 87–92.

    Article  PubMed  Google Scholar 

  24. Juárez Tomás, M. S., Bru, E., and Nader-Macías, M. E. (2002) Comparison of the growth and H2O2-production by vaginal probiotic lactobacilli under different culture conditions. Am. J. Obstet. Gynecol. 188, 35–44.

    Article  Google Scholar 

  25. Marty-Teysset, C., De La Torre, F., Garel, J. R. (2000) Increased production of hydrogen peroxide by Lactobacillus delbrueckii subsp. bulgaricus upon aeration: Involvement of an NADH oxidase in oxidative stress. Appl. Env. Microbiol. 66, 262–267.

    Article  CAS  Google Scholar 

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© 2004 Humana Press Inc. Totowa, NJ

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Tomás, M.S.J., Otero, M.C., Ocaña, V., Nader-Macías, M.E. (2004). Production of Antimicrobial Substances by Lactic Acid Bacteria I. In: Spencer, J.F.T., Ragout de Spencer, A.L. (eds) Public Health Microbiology. Methods in Molecular Biology, vol 268. Humana Press. https://doi.org/10.1385/1-59259-766-1:337

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  • DOI: https://doi.org/10.1385/1-59259-766-1:337

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-117-2

  • Online ISBN: 978-1-59259-766-6

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