Skip to main content

Advertisement

SpringerLink
Log in

Identification of Vaginal Lactobacilli with Potential Probiotic Properties Isolated from Women in North Lebanon

  • Host Microbe Interactions
  • Published:
Microbial Ecology Aims and scope Submit manuscript

Abstract

The aim of this work was to study the diversity of vaginal lactobacilli in Lebanese women and to evaluate the antagonism, hydrophobicity, and safety characteristics of these strains. This study was performed on samples from 135 women who visited a gynecology clinic in the north of Lebanon, between September 2012 and January 2013. From these samples, 53 different isolates of vaginal lactobacilli were collected from vaginal swabs and identified using biochemical and molecular methods. The use of genotypic Rep-PCR fingerprinting allowed for the organization of these isolates into 23 different groups. Seven of the isolated lactobacilli were antagonistic against the following vaginal pathogens: Gardnerella vaginalis CIP7074T, Staphylococcus aureus ATCC33862, Escherichia coli CIP103982, and Candida albicans ATCC10231. The antagonistic lactobacilli strains were then identified using 16S rDNA sequence. The data of this study show that the antagonistic lactobacilli were non-hemolytic, sensitive to most antibiotic tests, free of plasmid DNA, and exhibited interesting hydrophobicity and autoaggregation properties positioning them as potential candidates for probiotic design.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Andrade MJ, Rodriguez M, Sánchez B, Aranda E, Córdoba JJ (2006) DNA typing methods for differentiation of yeasts related to dry-cured meat products. Int J Food Microbiol 107:48–58

    Article  CAS  PubMed  Google Scholar 

  2. Antonio MA, Rabe LK, Hillier S (2005) Colonization of the rectum by Lactobacillus species and decreased risk of bacterial vaginosis. J Infect Dis 192:394–398

    Article  PubMed  Google Scholar 

  3. Antonio MA, Hawes SE, Hillier SL (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

    Article  CAS  PubMed  Google Scholar 

  4. Aslim B, Onal D, Beyatli Y (2007) Factors influencing autoaggregation and aggregation of Lactobacillus delbrueckii subsp. bulgaricus isolated from handmade yogurt. J Food Prot 70:223–227

    PubMed  Google Scholar 

  5. Batdorj B, Dalgalarrondo M, Choiset Y, Pedroche J, Métro F, Prévost H, Chobert JM, Haertlé T (2006) Purification and characterization of two bacteriocins produced by lactic acid bacteria isolated from Mongolian airag. J Appl Microbiol 101:837–848

    Article  CAS  PubMed  Google Scholar 

  6. Begovic J, Huys G, Mayo B, D’Haene K, Florez AB, Lozo J, Kojic M, Strahinic I, Topisirovic L (2009) Human vaginal Lactobacillus rhamnosus harbor mutation in 23S rRNA associated with erythromycin resistance. Res Microbiol 160:421–426

    Article  CAS  PubMed  Google Scholar 

  7. Belletti N, Gatti M, Bottari B, Neviani E, Tabanelli G, Gardini F (2009) Antibiotic resistance of lactobacilli isolated from two Italian hard cheeses. J Food Prot 72:2162–2169

    CAS  PubMed  Google Scholar 

  8. Bellon-fontaine MN, Rault J, van Oss CJ (1996) Microbial adhesion to solvents: a novel method to determine the electron-donor/electron-acceptor or Lewis acid-base properties of microbial cells. Colloids Surf B: Biointerfaces 7:47–53

    Article  CAS  Google Scholar 

  9. Boris S, Suárez JE, Vázquez F, Barbés C (1998) Adherence of human vaginal lactobacilli to vaginal epithelial cells and interaction with uropathogens. Infect Immun 66:1985–1989

    PubMed Central  CAS  PubMed  Google Scholar 

  10. Brotman RM, Erbelding EJ, Jamshidi RM, Klebanoff MA, Zenilman JM, Ghanem KG (2007) Findings associated with recurrence of bacterial vaginosis among adolescents attending sexually transmitted diseases clinics. J Pediatr Adolesc Gynecol 20:225–231

    Article  PubMed Central  PubMed  Google Scholar 

  11. Del Re B, Sgorbati B, Miglioli M, Palenzona D (2000) Adhesion, autoaggregation and hydrophobicity of 13 strains of Bifidobacterium longum. Lett Appl Microbiol 31:438–442

    Article  PubMed  Google Scholar 

  12. de Man JC, Rogosa M, Sharpe ME (1960) A medium for the cultivation of lactobacilli. J Appl Bacteriol 23:130–135

    Article  Google Scholar 

  13. Dimitonova SP, Bakalov BV, Aleksandrova-Georgieva RN, Danova ST (2008) Phenotypic and molecular identification of lactobacilli isolated from vaginal secretions. J Microbiol Immunol Infect 41:469–477

    CAS  PubMed  Google Scholar 

  14. Drago L, Mattina R, Nicola L, Rodighiero V, De Vechi E (2011) Macrolide resistance and in vitro selection of resistance to antibiotics in Lactobacillus isolates. J Microbiol 49:651–656

    Article  CAS  PubMed  Google Scholar 

  15. Dubernet S, Desmasures N, Guéguen M (2002) A PCR-based method for identification of lactobacilli at the genus level. FEMS Microbiol Lett 21:271–275

    Article  Google Scholar 

  16. Dunne C, O’Mahony L, Thornton G, Morrissey D, O’Halloran S, Feeney M, Flynn S, Fitzgerald G, Daly C, Kiely B, O’Sullivan GC, Shanahan F, Collins JK (2001) In vitro selection criteria for probiotic bacteria of human origin: correlation with in vivo findings. Am J Clin Nutr 73:386–392

    Google Scholar 

  17. FAO/WHO (2006) Probiotics in food. Health and nutritional properties and guidelines for evaluation. FAO Food and Nutritional paper No. 85 (ISBN 92-5-105513-0)

  18. Felten A, Barreau C, Bizet C, Lagrange PH, Philippon A (1999) Lactobacillus species identification, H2O2 and antibiotic resistance and correlation with human clinical status. J Clin Microbiol 37:729–733

    PubMed Central  CAS  PubMed  Google Scholar 

  19. Fichorova RN, Onderdonk AB, Yamamoto H, Delaney ML, Dubois AM, Allred E, Leviton A (2011) Maternal microbe-specific modulation of inflammatory response in extremely low-gestational-age newborns. MBio 2:280–10

    Google Scholar 

  20. Garg KB, Ganguli I, Das R, Talwar GP (2009) Spectrum of Lactobacillus species present in healthy vagina of Indian women. Indian J Med Res 129:652–657

    CAS  PubMed  Google Scholar 

  21. Gil NF, Martinez RC, Gomes BC, Nomizo A, De Martinis EC (2010) Vaginal lactobacilli as potential probiotics against Candida spp. Braz J Microbiol 41:6–14

    Article  PubMed  Google Scholar 

  22. Greene JD, Klaenhammer TR (1994) Factors involved in adherence of lactobacilli to human Caco-2 cells. Appl Environ Microbiol 60:4487–4494

    PubMed Central  CAS  PubMed  Google Scholar 

  23. Hsieh PS, Tsai YC, Chen YC, Teh SF, Ou CM, King VA (2012) Eradication of Helicobacter pylori infection by the probiotic strains Lactobacillus johnsonii MH-68 and L. salivarius ssp. salicinius AP-32. Helicobacter 17:466–477

    Article  PubMed  Google Scholar 

  24. Hyman RW, Fukushima M, Jiang H, Fung E, Rand L, Johnson B, Vo KC, Caughey AB, Hilton JF, Davis RW, Giudice LC (2013) Diversity of the vaginal microbiome correlates with preterm birth. Reproductives Sciences 28

  25. Juarez Tomas MS, Zonenschain D, Morelli L, Nader Macias ME (2005) Characterization of potentially probiotic vaginal lactobacilli isolated from Argentinian women. Br J Biomed Sci 62:170–174

    CAS  PubMed  Google Scholar 

  26. Kabadjova P, Dousset X, Le Cam V, Prevost H (2002) Differentiation of closely related Carnobacterium food isolates based on 16S-23S ribosomal DNA intergenic spacer region polymorphism. Appl Environ Microbiol 68:5358–5366

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  27. Larsson PG, Bergstrom M, Forsum U, Jacobsson B, Strand A, Wolner-Hanssen P (2005) Bacterial vaginosis. Transmission, role in genital tract infection and pregnancy outcome: an enigma. APMIS 113:233–245

    Article  CAS  PubMed  Google Scholar 

  28. MacPhee RA, Miller WL, Gloor GB, McCormick JK, Hammond JA, Burton JP, Reid G (2013) Influence of the vaginal microbiota on toxic shock syndrome toxin 1 production by Staphylococcus aureus. Appl Environ Microbiol 79:1835–1842

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  29. Marcotte H, Ferrari S, Cesena C, Hammarstrom L, Morelli L, Pozzi G, Oggioni MR (2004) The aggregation-promoting factor of Lactobacillus crispatus M247 and its genetic locus. J Appl Microbiol 97:749–756

    Article  CAS  PubMed  Google Scholar 

  30. Martin HL, Richardson BA, Nyange PM, Lavreys L, Hillier SL, Chohan B, Mandaliya K, Ndinyaachola JO, Bwayo, Kreiss J (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  CAS  PubMed  Google Scholar 

  31. Martirosian G, Radosz-Komoniewska H, Pietrzak B, Ekiel A, Kaminski P, Aptekorz M, Dolezych H, Samulska E, Jozwiak J (2012) Characterization of vaginal lactobacilli after kidney transplantation. Anaerobe 18:209–213

    Article  CAS  PubMed  Google Scholar 

  32. Mastromarino P, Vitali B, Mosca L (2013) Bacterial vaginosis: a review on clinical trials with probiotics. New Microbiol 36:229–238

    PubMed  Google Scholar 

  33. Mastromarino P, Brigidi P, Macchia S, Maggi L, Pirovano F, Trinchieri V, Conte U, Matteuzzi D (2002) Characterization and selection of vaginal Lactobacillus strains for the preparation of vaginal tablets. J Appl Microbiol 93:884–893

    Article  CAS  PubMed  Google Scholar 

  34. Millsap K, Reid G, van der Mei C, Busscher H (1996) Cluster analysis of genotypically characterized Lactobacillus species based on physicochemical cell surfaces properties and their relationship with adhesion to hexdecane. Can J Microbiol 43:284–291

    Article  Google Scholar 

  35. Motevaseli E, Shirzad M, Raoofian R, Hasheminasab SM, Hatami M, Dianatpour M, Modarressi MH (2013) Differences in vaginal lactobacilli composition of Iranian healthy and bacterial vaginosis infected women: a comparative analysis of their cytotoxic effects with commercial vaginal probiotics. Iran Red Crescent Med J 15:199–206

    Article  PubMed Central  PubMed  Google Scholar 

  36. Newman D (1915) The treatment of cystitis by intravesical injections of lactic bacillus cultures. Lancet 330

  37. Nielsen DS, Schillinger U, Franz CM, Bresciani J, Amoa-Awua W, Holzapfel WH, Jakobsen M (2007) Lactobacillus ghanensis sp. nov., a motile lactic acid bacterium isolated from Ghanaian cocoa fermentations. Int J Syst Evol Microbiol 57:1468–1472

    Article  CAS  PubMed  Google Scholar 

  38. Nugent RP, Krohn MA, Hillier SL (1991) Reliability of diagnosing bacterial vaginosis is improved by a standardized method of gram stain interpretation. J Clin Microbiol 29:297–301

    PubMed Central  CAS  PubMed  Google Scholar 

  39. Osset J, Bartolomé RM, García E (2001) Assessment of the capacity of Lactobacillus to inhibit the growth of uropathogens and block their adhesion to vaginal epithelial cells. J Infect Dis 183:485–491

    Article  CAS  PubMed  Google Scholar 

  40. Pavlova SI, Kilic AO, Kilic SS, So JS, Nader-Macias ME, Simoes JA, 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

    Article  CAS  PubMed  Google Scholar 

  41. Pendharkar S, Magopane T, Larsson PG, de Bruyn G, Gray GE, Hammarström L, Marcotte H (2013) Identification and characterisation of vaginal lactobacilli from South African women. BMC Infect Dis 13:43

    Article  PubMed Central  PubMed  Google Scholar 

  42. Pelletier C, Bouley C, Cayuela C, Bouttier S, Bourlioux P, Bellon-Fontaine MN (1997) Cell surface characteristics of Lactobacillus casei subsp. casei, Lactobacillus paracasei subsp. paracasei, and Lactobacillus rhamnosus strains. Appl Environ Microbiol 63:1725–1731

    PubMed Central  CAS  PubMed  Google Scholar 

  43. Petrova MI, van den Broek M, Balzarini J, Vanderleyden J, Lebeer S (2013) Vaginal microbiota and its role in HIV transmission and infection. FEMS Microbiol Rev 7:762–792

    Article  Google Scholar 

  44. Reid G (2012) Probiotic and prebiotic applications for vaginal health. J AOAC Int 295:31–34

    Article  Google Scholar 

  45. Rojas M, Conway PL (1996) Colonization by lactobacilli of piglet small intestinal mucus. J Appl Microbiol 81:474–480

    Article  CAS  Google Scholar 

  46. Rosenberg M, Gutnick D, Rosenberg E (1980) Adherence of bacteria to hydrocarbons: a simple method for measuring cell-surface hydrophobicity. FEMS Microbiol Lett 9:29–33

    Article  CAS  Google Scholar 

  47. Ruiz FO, Gerbaldo G, Garcia MJ, Giordano W, Pascual L, Barberis L (2012) Synergistic effect between two bacteriocin-like substances produced by lactobacilli strains with inhibitory activity for Streptococcus agalactiae. Curr Microbiol 64:349–356

    Article  CAS  PubMed  Google Scholar 

  48. Sánchez B, González-Tejedo C, Ruas-Madiedo P, Urdaci MC, Margolles A (2010) Lactobacillus plantarum extracellular chitin-binding protein and its role in the interaction between chitin, Caco-2 cells, and mucin. Appl Environ Microbiol 77:1123–1126

    Article  PubMed Central  PubMed  Google Scholar 

  49. Sewankambo N, Gray RH, Wawer MJ, Paxton L, McNaim D, Wabwire-Mangen F, Serwadda D, Li C, Kiwanuka N, Hillier SL, Rabe L, Gaydos CA, Quinn TC, Konde-Lule J (1997) HIV-1 infection associated with abnormal vaginal flora morphology and bacterial vaginosis. Lancet 23:546–550, Erratum in: Lancet 1997, 350:1036

    Article  Google Scholar 

  50. Sivester ME, Dicks LM (2003) identification of lactic acid bacteria from human vaginal secretions. Antonie Van Leeuwenhoek 83:117–123

    Article  Google Scholar 

  51. Song YL, Kato N, Matsumiya Y, Liu CX, Kato H, Watanabe K (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

    PubMed Central  CAS  PubMed  Google Scholar 

  52. Steen A, Buist G, Leenhouts KJ, El Khattabi M, Grijpstra F, Zomer AL, Venema G, Kuipers OP, Kok J (2003) Cell wall attachment of a widely distributed peptidoglycan binding domain is hindered by cell wall constituents. J Biol Chem 278:23874–23881

    Article  CAS  PubMed  Google Scholar 

  53. Tagg JR, McGiven AR (1971) Assay system for bacteriocins. Appl Microbiol 21:943

    PubMed Central  CAS  PubMed  Google Scholar 

  54. Tarnberg M, Jakobsson T, Jonasson J, Forsum U (2002) Identification of randomly selected colonies of lactobacilli from normal vaginal fluid by pyrosequencing of the 16S rRNA variable V1 and V3 regions. Acta Pathol Microbiol Immunol Scand 110:802–810

    Article  Google Scholar 

  55. Tulini FL, Winkelströter LK, De Martinis ECP (2013) Identification and evaluation of the probiotic potential of Lactobacillus paraplantarum FT259, a bacteriocinogenic strain isolated from Brazilian semi-hard artisanal cheese. Anaerobe 22:57–63

    Article  CAS  PubMed  Google Scholar 

  56. Vásquez A, Jakobsson T, Ahrné S, Forsum U, Molin G (2002) Vaginal Lactobacillus flora of healthy Swedish women. J Clin Microbiol 40:2746–2749

    Article  PubMed Central  PubMed  Google Scholar 

  57. Wilks M, Wiggins R, Whiley A, Hennessy E, Warwick S, Porter H, Corfield A, Millar M (2004) Identification and H2O2 production of vaginal lactobacilli from pregnant women at high risk of preterm birth and relation with outcome. J Clin Microbiol 42:713–717

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  58. Yuksekdag ZN, Beyatli Y (2008) Antimicrobial activity and antibiotic susceptibility of some lactic acid bacteria isolated from different dairy products. Arch Leb 59:216–220

    CAS  Google Scholar 

  59. Younes JA, van der Mai HC, van den Heuvel E, Busscher HJ, Reid G (2012) Adhesion forces and coaggregation between vaginal staphylococcal and lactobacilli. PLoS ONE 7:e36917

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  60. 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 1:121–133

    Article  CAS  PubMed  Google Scholar 

  61. Zhou JS, Pillidge CJ, Gopal PK, Gill HS (2005) Antibiotic susceptibility profiles of new probiotic Lactobacillus and Bifidobacterium strains. Int J Food Microbiol 98:211–217

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was possible through the financial support of Lebanese University and Lille1 University. The authors would like to thank Prof. Mike Chikindas (Rutgers University) and Delphine Caly (Lille 1 University) for the critical reading of the manuscript. The authors are indebted to Mr. Taha ABDOU and Mrs Mariam YEHIA for their excellent technical assistance.

Author information

Authors and Affiliations

  1. Laboratoire des Procédés Biologiques, Génie Enzymatique et Microbien (ProBioGEM), Cité Scientifique, avenue Paul Langevin. Bâtiment—Polytech Lille, 59655, Villeneuve d’Ascq Cedex, France

    Imad Al Kassaa, Nour-Eddine Chihib & Djamel Drider

  2. Laboratoire de Microbiologie Santé et Environnement, Rue Mitein, Centre AZM de Biotechnologie, EDST-Université Libanaise, Tripoli, Lebanon

    Imad Al Kassaa & Monzer Hamze

  3. Faculté de Santé Publique Section 3, Université Libanaise, Tripoli, Lebanon

    Monzer Hamze

  4. Laboratoire de Virologie EA3610 Université Lille 2, CHRU Lille, 59120, Loos-lez-Lille, France

    Didier Hober

Authors
  1. Imad Al Kassaa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Monzer Hamze
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Didier Hober
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nour-Eddine Chihib
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Djamel Drider
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Djamel Drider.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Al Kassaa, I., Hamze, M., Hober, D. et al. Identification of Vaginal Lactobacilli with Potential Probiotic Properties Isolated from Women in North Lebanon. Microb Ecol 67, 722–734 (2014). https://doi.org/10.1007/s00248-014-0384-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00248-014-0384-7

Keywords

Search

Navigation