1887

Abstract

Within the vaginal ecosystem, lactobacilli and spp. likely interact and influence each other’s growth, yet the details of this interaction are not clearly defined. Using medium simulating vaginal fluid and a two-chamber co-culturing system to prevent cell-to-cell contact between the bacteria, we examined the possibility that 62B (Lj 62B) and/or (Gp) JCP8151B produce extracellular factors through which they influence each other’s viability. By 24 h post-inoculation (hpi) in the co-culture system and under conditions similar to the vaginal environment – pH 5.0, 37 °C, and 5% CO, Lj 62B viability was not affected but Gp JCP8151B had been eliminated. Cell-free supernatant harvested from Lj 62B cultures (Lj-CFS) at 20 hpi, but not 16 hpi, also eliminated Gp JCP8151B growth. Neither lactic acid nor HO production by Lj 62B was responsible for this effect. The Lj-CFS did not affect viability of three species of lactobacilli or eight species of Gram-positive and Gram-negative uropathogens but eliminated viability of eight different strains of spp. Activity of the inhibitory factor within Lj-CFS was abolished by protease treatment and reduced by heat treatment suggesting it is most likely a bacteriocin-like protein; fractionation revealed that the factor has a molecular weight within the 10–30 kDa range. These results suggest that, in medium mimicking vaginal fluid and growth conditions similar to the vaginal environment, Lj 62B produces a potential bacteriocin-like inhibitory substance (Lj-BLIS) that clearly targets spp. strains. Once fully characterized, Lj-BLIS may be a potential treatment for related BV that does not alter the vaginal microflora.

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2023-11-01
2024-05-10
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References

  1. Chen X, Lu Y, Chen T, Li R. The female vaginal microbiome in health and bacterial vaginosis. Front Cell Infect Microbiol 2021; 11:631972 [View Article] [PubMed]
    [Google Scholar]
  2. Ravel J, Gajer P, Abdo Z, Schneider GM, Koenig SSK et al. Vaginal microbiome of reproductive-age women. Proc Natl Acad Sci U S A 2011; 108 Suppl 1:4680–4687 [View Article] [PubMed]
    [Google Scholar]
  3. Witkin SS, Linhares IM. Why do lactobacilli dominate the human vaginal microbiota?. BJOG 2017; 124:606–611 [View Article] [PubMed]
    [Google Scholar]
  4. Miller EA, Beasley DE, Dunn RR, Archie EA. Lactobacilli dominance and vaginal pH: why is the human vaginal microbiome unique?. Front Microbiol 2016; 7:1936 [View Article] [PubMed]
    [Google Scholar]
  5. Pavlova SI, Kilic AO, Kilic SS, So J-S, Nader-Macias ME et al. Genetic diversity of vaginal lactobacilli from women in different countries based on 16S rRNA gene sequences. J Appl Microbiol 2002; 92:451–459 [View Article] [PubMed]
    [Google Scholar]
  6. Vásquez A, Jakobsson T, Ahrné S, Forsum U, Molin G. Vaginal lactobacillus flora of healthy Swedish women. J Clin Microbiol 2002; 40:2746–2749 [View Article] [PubMed]
    [Google Scholar]
  7. Boskey ER, Cone RA, Whaley KJ, Moench TR. Origins of vaginal acidity: high D/L lactate ratio is consistent with bacteria being the primary source. Hum Reprod 2001; 16:1809–1813 [View Article] [PubMed]
    [Google Scholar]
  8. Juárez Tomás MS, Ocaña VS, Wiese B, Nader-Macías ME. Growth and lactic acid production by vaginal Lactobacillus acidophilus CRL 1259, and inhibition of uropathogenic Escherichia coli. J Med Microbiol 2003; 52:1117–1124 [View Article] [PubMed]
    [Google Scholar]
  9. O’Hanlon DE, Come RA, Moench TR. Vaginal pH measured in vivo: lactobacilli determine pH and lactic acid concentration. BMC Microbiol 2019; 19:13 [View Article] [PubMed]
    [Google Scholar]
  10. O’Hanlon DE, Moench TR, Cone RA. In vaginal fluid, bacteria associated with bacterial vaginosis can be suppressed with lactic acid but not hydrogen peroxide. BMC Infect Dis 2011; 11:200 [View Article] [PubMed]
    [Google Scholar]
  11. Atassi F, Servin AL. Individual and co-operative roles of lactic acid and hydrogen peroxide in the killing activity of enteric strain Lactobacillus johnsonii NCC933 and vaginal strain Lactobacillus gasseri KS120.1 against enteric, uropathogenic and vaginosis-associated pathog. FEMS Microbiol Lett 2010; 304:2938 [View Article]
    [Google Scholar]
  12. Hawes SE, Hillier SL, Benedetti J, Stevens CE, Koutsky LA et al. Hydrogen peroxide-producing lactobacilli and acquisition of vaginal infections. J Infect Dis 1996; 174:1058–1063 [View Article] [PubMed]
    [Google Scholar]
  13. Kovachev S. Defence factors of vaginal lactobacilli. Crit Rev Microbiol 2018; 44:31–39 [View Article] [PubMed]
    [Google Scholar]
  14. Stoyancheva G, Marzotto M, Dellaglio F, Torriani S. Bacteriocin production and gene sequencing analysis from vaginal Lactobacillus strains. Arch Microbiol 2014; 196:645–653 [View Article] [PubMed]
    [Google Scholar]
  15. Gradisteanu Pircalabioru G, Popa LI, Marutescu L, Gheorghe I, Popa M et al. Bacteriocins in the era of antibiotic resistance: rising to the challenge. Pharmaceutics 2021; 13:196 [View Article]
    [Google Scholar]
  16. Alvarez-Sieiro P, Montalbán-López M, Mu D, Kuipers OP. Bacteriocins of lactic acid bacteria: extending the family. Appl Microbiol Biotechnol 2016; 100:2939–2951 [View Article] [PubMed]
    [Google Scholar]
  17. Hernández-González JC, Martínez-Tapia A, Lazcano-Hernández G, García-Pérez BE, Castrejón-Jiménez NS. Bacteriocins from lactic acid bacteria. A powerful alternative as antimicrobials, probiotics, and immunomodulators in veterinary medicine. Animals 2021; 11:979 [View Article] [PubMed]
    [Google Scholar]
  18. Pérez-Ramos A, Madi-Moussa D, Coucheney F, Drider D. Current knowledge of the mode of action and immunity mechanisms of LAB-bacteriocins. Microorganisms 2021; 9:2107 [View Article] [PubMed]
    [Google Scholar]
  19. Aranha C, Gupta S, Reddy KVR. Contraceptive efficacy of antimicrobial peptide Nisin: in vitro and in vivo studies. Contraception 2004; 69:333–338 [View Article] [PubMed]
    [Google Scholar]
  20. Dover SE, Aroutcheva AA, Faro S, Chikindas ML. Safety study of an antimicrobial peptide lactocin 160, produced by the vaginal Lactobacillus rhamnosus. Infect Dis Obstet Gynecol 2007; 2007:78248 [View Article] [PubMed]
    [Google Scholar]
  21. Kairys N, Garg M. Bacterial Vaginosis. Treasure Island (FL): StatPearls; 2022
  22. Ravel J, Moreno I, Simón C. Bacterial vaginosis and its association with infertility, endometritis, and pelvic inflammatory disease. Am J Obstet Gynecol 2021; 224:251–257 [View Article] [PubMed]
    [Google Scholar]
  23. Janulaitiene M, Paliulyte V, Grinceviciene S, Zakareviciene J, Vladisauskiene A et al. Prevalence and distribution of Gardnerella vaginalis subgroups in women with and without bacterial vaginosis. BMC Infect Dis 2017; 17:394 [View Article] [PubMed]
    [Google Scholar]
  24. Donders GGG. Definition and classification of abnormal vaginal flora. Best Pract Res Clin Obstet Gynaecol 2007; 21:355–373 [View Article] [PubMed]
    [Google Scholar]
  25. Martin DH, Marrazzo JM. The vaginal microbiome: current understanding and future directions. J Infect Dis 2016; 214 Suppl 1:S36–41 [View Article] [PubMed]
    [Google Scholar]
  26. Onderdonk AB, Delaney ML, Fichorova RN. The human microbiome during bacterial vaginosis. Clin Microbiol Rev 2016; 29:223–238 [View Article] [PubMed]
    [Google Scholar]
  27. Hill JE, Albert AYK. VOGUE Research Group Resolution and cooccurrence patterns of Gardnerella leopoldii, G. swidsinskii, G. piotii, and G. vaginalis within the vaginal microbiome. Infect Immun 2019; 87:e00532-19 [View Article] [PubMed]
    [Google Scholar]
  28. Khan S, Voordouw MJ, Hill JE. Competition among Gardnerella subgroups from the human vaginal microbiome. Front Cell Infect Microbiol 2019; 9:374 [View Article] [PubMed]
    [Google Scholar]
  29. Vaneechoutte M, Guschin A, Van Simaey L, Gansemans Y, Van Nieuwerburgh F et al. Emended description of Gardnerella vaginalis and description of Gardnerella leopoldii sp. nov., Gardnerella piotii sp. nov. and Gardnerella swidsinskii sp. nov., with delineation of 13 genomic species within the genus Gardnerella. Int J Syst Evol Microbiol 2019; 69:679–687 [View Article] [PubMed]
    [Google Scholar]
  30. Qin H, Xiao B. Research progress on the correlation between Gardnerella typing and bacterial vaginosis. Front Cell Infect Microbiol 2022; 12:858155 [View Article] [PubMed]
    [Google Scholar]
  31. Pybus V, Onderdonk AB. Microbial interactions in the vaginal ecosystem, with emphasis on the pathogenesis of bacterial vaginosis. Microbes Infect 1999; 1:285–292 [View Article] [PubMed]
    [Google Scholar]
  32. Valore EV, Wiley DJ, Ganz T. Reversible deficiency of antimicrobial polypeptides in bacterial vaginosis. Infect Immun 2006; 74:56935702 [View Article] [PubMed]
    [Google Scholar]
  33. Haggerty CL, Hillier SL, Bass DC, Ness RB, Evaluation PID. PID Evaluation and Clinical Health study investigators Bacterial vaginosis and anaerobic bacteria are associated with endometritis. Clin Infect Dis 2004; 39:990–995 [View Article] [PubMed]
    [Google Scholar]
  34. Mirmonsef P, Spear GT. The barrier to HIV transmission provided by genital tract Lactobacillus colonization. Am J Reprod Immunol 2014; 71:531–536 [View Article] [PubMed]
    [Google Scholar]
  35. Schwebke JR, Desmond R. Risk factors for bacterial vaginosis in women at high risk for sexually transmitted diseases. Sex Transm Dis 2005; 32:654-8 [View Article] [PubMed]
    [Google Scholar]
  36. Wiesenfeld HC, Hillier SL, Krohn MA, Amortegui AJ, Heine RP et al. Lower genital tract infection and endometritis: insight into subclinical pelvic inflammatory disease. Obstet Gynecol 2002; 100:456–463 [View Article] [PubMed]
    [Google Scholar]
  37. Cauci S, Culhane JF, Di Santolo M, McCollum K. Among pregnant women with bacterial vaginosis, the hydrolytic enzymes sialidase and prolidase are positively associated with interleukin-1beta. Am J Obstet Gynecol 2008; 198:132 [View Article] [PubMed]
    [Google Scholar]
  38. Udayalaxmi J, Bhat GK, Kotigadde S. Biotypes and virulence factors of Gardnerella vaginalis isolated from cases of bacterial vaginosis. Indian J Med Microbiol 2011; 29:165–168 [View Article] [PubMed]
    [Google Scholar]
  39. Yeoman CJ, Yildirim S, Thomas SM, Durkin AS, Torralba M et al. Comparative genomics of Gardnerella vaginalis strains reveals substantial differences in metabolic and virulence potential. PLoS One 2010; 5:e12411 [View Article] [PubMed]
    [Google Scholar]
  40. Bradshaw CS, Pirotta M, De Guingand D, Hocking JS, Morton AN et al. Efficacy of oral metronidazole with vaginal clindamycin or vaginal probiotic for bacterial vaginosis: randomised placebo-controlled double-blind trial. PLoS One 2012; 7:e34540 [View Article] [PubMed]
    [Google Scholar]
  41. Eriksson K, Carlsson B, Forsum U, Larsson P-G. A double-blind treatment study of bacterial vaginosis with normal vaginal lactobacilli after an open treatment with vaginal clindamycin ovules. Acta Derm Venereol 2005; 85:42–46 [View Article] [PubMed]
    [Google Scholar]
  42. Paavonen J, Mangioni C, Martin MA, Wajszczuk CP. Vaginal clindamycin and oral metronidazole for bacterial vaginosis: a randomized trial. Obstet Gynecol 2000; 96:256–260 [View Article] [PubMed]
    [Google Scholar]
  43. Karpinets TV, Solley TN, Mikkelson MD, Dorta-Estremera S, Nookala SS et al. Effect of antibiotics on gut and vaginal microbiomes associated with cervical cancer development in mice. Cancer Prev Res 2020; 13:997–1006 [View Article] [PubMed]
    [Google Scholar]
  44. van de Wijgert J, Verwijs MC. Lactobacilli-containing vaginal probiotics to cure or prevent bacterial or fungal vaginal dysbiosis: a systematic review and recommendations for future trial designs. BJOG 2020; 127:287–299 [View Article] [PubMed]
    [Google Scholar]
  45. van de Wijgert JHHM, Verwijs MC, Agaba SK, Bronowski C, Mwambarangwe L et al. Intermittent lactobacilli-containing vaginal probiotic or metronidazole use to prevent bacterial vaginosis recurrence: A pilot study incorporating microscopy and sequencing. Sci Rep 2020; 10:3884 [View Article] [PubMed]
    [Google Scholar]
  46. Katkowska M, Garbacz K, Kusiak A. Probiotics: should all patients take them?. Microorganisms 2021; 9:2620 [View Article] [PubMed]
    [Google Scholar]
  47. Rossi F, Amadoro C, Gasperi M, Colavita G. Lactobacilli infection case reports in the last three years and safety implications. Nutrients 2022; 14:1178 [View Article] [PubMed]
    [Google Scholar]
  48. Valenti P, Rosa L, Capobianco D, Lepanto MS, Schiavi E et al. Role of lactobacilli and lactoferrin in the mucosal cervicovaginal defense. Front Immunol 2018; 9:376 [View Article] [PubMed]
    [Google Scholar]
  49. Wang S, Wang Q, Yang E, Yan L, Li T et al. Antimicrobial compounds produced by vaginal Lactobacillus crispatus are able to strongly inhibit Candida albicans growth, hyphal formation and regulate virulence-related gene expressions. Front Microbiol 2017; 8:564 [View Article] [PubMed]
    [Google Scholar]
  50. Juárez Tomás MS, Nader-Macías MEF. Effect of a medium simulating vaginal fluid on the growth and expression of beneficial characteristics of potentially probiotic lactobacilli. In Méndez-Vilas A. eds Communicating Current Research and Educational Topics and Trends in Applied Microbiology vol 2 Badajoz, Spain: FORMATEX; 2007 pp 732–739
    [Google Scholar]
  51. Navarro S, Abla H, Delgado B, Colmer-Hamood JA, Ventolini G et al. Glycogen availability and pH variation in a medium simulating vaginal fluid influence the growth of vaginal Lactobacillus species and Gardnerella vaginalis. BMC Microbiol 2023; 23:186 [View Article] [PubMed]
    [Google Scholar]
  52. Putonti C, Thomas-White K, Crum E, Hilt EE, Price TK et al. Genome investigation of urinary Gardnerella strains and their relationship to isolates of the vaginal microbiota. mSphere 2021; 6:e00154-21 [View Article] [PubMed]
    [Google Scholar]
  53. Mitchell C, Fredricks D, Agnew K, Hitti J. Hydrogen peroxide-producing lactobacilli are associated with lower levels of vaginal interleukin-1beta, independent of bacterial vaginosis. Sex Transm Dis 2015; 42:358–363 [View Article] [PubMed]
    [Google Scholar]
  54. Tachedjian G, Aldunate M, Bradshaw CS, Cone RA. The role of lactic acid production by probiotic Lactobacillus species in vaginal health. Res Microbiol 2017; 168:782792 [View Article] [PubMed]
    [Google Scholar]
  55. Maldonado-Barragán A, Caballero-Guerrero B, Martín V, Ruiz-Barba JL, Rodríguez JM. Purification and genetic characterization of gassericin E, a novel co-culture inducible bacteriocin from Lactobacillus gasseri EV1461 isolated from the vagina of a healthy woman. BMC Microbiol 2016; 16:37 [View Article] [PubMed]
    [Google Scholar]
  56. Holo H, Nilssen O, Nes IF. Lactococcin A, a new bacteriocin from Lactococcus lactis subsp. cremoris: isolation and characterization of the protein and its gene. J Bacteriol 1991; 173:3879–3887 [View Article] [PubMed]
    [Google Scholar]
  57. Martínez B, Suárez JE, Rodríguez A. Lactococcin 972 : a homodimeric lactococcal bacteriocin whose primary target is not the plasma membrane. Microbiology 1996; 142 (Pt 9):2393–2398 [View Article] [PubMed]
    [Google Scholar]
  58. Venema K, Abee T, Haandrikman AJ, Leenhouts KJ, Kok J et al. Mode of action of lactococcin b, a thiol-activated bacteriocin from Lactococcus lactis. Appl Environ Microbiol 1993; 59:1041–1048 [View Article] [PubMed]
    [Google Scholar]
  59. Rea MC, Dobson A, O’Sullivan O, Crispie F, Fouhy F et al. Effect of broad- and narrow-spectrum antimicrobials on Clostridium difficile and microbial diversity in a model of the distal colon. Proc Natl Acad Sci U S A 2011; 108 Suppl 1:4639–4644 [View Article] [PubMed]
    [Google Scholar]
  60. Corr SC, Li Y, Riedel CU, O’Toole PW, Hill C et al. Bacteriocin production as a mechanism for the antiinfective activity of Lactobacillus salivarius UCC118. Proc Natl Acad Sci U S A 2007; 104:7617–7621 [View Article] [PubMed]
    [Google Scholar]
  61. Kaewsrichan J, Peeyananjarassri K, Kongprasertkit J. Selection and identification of anaerobic lactobacilli producing inhibitory compounds against vaginal pathogens. FEMS Immunol Med Microbiol 2006; 48:75–83 [View Article] [PubMed]
    [Google Scholar]
  62. Matu MN, Orinda GO, Njagi ENM, Cohen CR, Bukusi EA. In vitro inhibitory activity of human vaginal lactobacilli against pathogenic bacteria associated with bacterial vaginosis in Kenyan women. Anaerobe 2010; 16:210–215 [View Article] [PubMed]
    [Google Scholar]
  63. Aroutcheva AA, Simoes JA, Faro S. Antimicrobial protein produced by vaginal Lactobacillus acidophilus that inhibits Gardnerella vaginalis. Infect Dis Obstet Gynecol 2001; 9:33–39 [View Article] [PubMed]
    [Google Scholar]
  64. Beukes M, Bierbaum G, Sahl HG, Hastings JW. Purification and partial characterization of a murein hydrolase, millericin B, produced by Streptococcus milleri NMSCC 061. Appl Environ Microbiol 2000; 66:23–28 [View Article] [PubMed]
    [Google Scholar]
  65. Swe PM, Cook GM, Tagg JR, Jack RW. Mode of action of dysgalacticin: a large heat-labile bacteriocin. J Antimicrob Chemother 2009; 63:679–686 [View Article] [PubMed]
    [Google Scholar]
  66. Ocaña VS, Pesce De Ruiz Holgado AA, Nader-Macías ME. Characterization of a bacteriocin-like substance produced by a vaginal Lactobacillus salivarius strain. Appl Environ Microbiol 1999; 65:5631–5635 [View Article] [PubMed]
    [Google Scholar]
  67. Goh HF, Philip K. Purification and characterization of bacteriocin produced by Weissella confusa A3 of dairy origin. PLoS One 2015; 10:e0140434 [View Article] [PubMed]
    [Google Scholar]
  68. Mahrous H, Mohamed A, El-Mongy MA, El-Batal AI, Hamza HA. Study bacteriocin production and optimization using new isolates of Lactobacillus spp. isolated from some dairy products under different culture conditions. Food Nutr Sci 2013; 04:342–356 [View Article]
    [Google Scholar]
  69. Dai M, Li Y, Xu L, Wu D, Zhou Q et al. A novel bacteriocin from Lactobacillus pentosus ZFM94 and its antibacterial mode of action. Front Nutr 2021; 8:710862 [View Article] [PubMed]
    [Google Scholar]
  70. Gaspar C, Donders GG, Palmeira-de-Oliveira R, Queiroz JA, Tomaz C et al. Bacteriocin production of the probiotic Lactobacillus acidophilus KS400. AMB Express 2018; 8:153 [View Article] [PubMed]
    [Google Scholar]
  71. Jawan R, Abbasiliasi S, Tan JS, Mustafa S, Halim M et al. Influence of culture conditions and medium compositions on the production of bacteriocin-like inhibitory substances by Lactococcus lactis Gh1. Microorganisms 2020; 8:1454 [View Article]
    [Google Scholar]
  72. Moon EC, Park MS, Lim T, Kim RH, Ji GE et al. Antibacterial effect of cell-free supernatant fraction from Lactobacillus paracasei CH88 against Gardnerella vaginalis. Sci Rep 2022; 12:4763 [View Article] [PubMed]
    [Google Scholar]
  73. Navarro S, Sherman E, Colmer-Hamood JA, Nelius T, Myntti M et al. Urinary catheters coated with a novel biofilm preventative agent inhibit biofilm development by diverse bacterial uropathogens. Antibiotics 2022; 11:1514 [View Article] [PubMed]
    [Google Scholar]
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