1887

Journal of Medical Microbiology logo

Volume 73, Issue 6

The effect of daily usage of Listerine Cool Mint mouthwash on the oropharyngeal microbiome: a substudy of the PReGo trial No Access

Abstract

Listerine is a bactericidal mouthwash widely used to prevent oral health problems such as dental plaque and gingivitis. However, whether it promotes or undermines a healthy oral microbiome is unclear.

We hypothesized that the daily use of Listerine Cool Mint would have a significant impact on the oropharyngeal microbiome.

We aimed to assess if daily usage of Listerine Cool Mint influenced the composition of the pharyngeal microbiome.

The current microbiome substudy is part of the Preventing Resistance in Gonorrhoea trial. This was a double-blind single-centre, crossover, randomized controlled trial of antibacterial versus placebo mouthwash to reduce the incidence of gonorrhoea/chlamydia/syphilis in men who have sex with men (MSM) taking HIV pre-exposure prophylaxis (PrEP). Fifty-nine MSM taking HIV PrEP were enrolled. In this crossover trial, participants received 3 months of daily Listerine followed by 3 months of placebo mouthwash or vice versa. Oropharyngeal swabs were taken at baseline and after 3 months use of each mouthwash. DNA was extracted for shotgun metagenomic sequencing (Illumina Inc.). Non-host reads were taxonomically classified with MiniKraken and Bracken. The alpha and beta diversity indices were compared between baseline and after each mouthwash use. Differentially abundant bacterial taxa were identified using ANOVA-like differential expression analysis.

was the most abundant genus in most samples ( = 103, 61.7 %) with a median relative abundance of 31.5% (IQR 20.6–44.8), followed by [13.5% (IQR 4.8–22.6)] and [10.0% (IQR 4.0–16.8)]. Compared to baseline, the composition of the oral microbiome at the genus level (beta diversity) was significantly different after 3 months of Listerine ( = 0.006, pseudo- = 2.29) or placebo ( = 0.003, pseudo- = 2.49, permutational multivariate analysis of variance) use. and were significantly more abundant after Listerine use compared to baseline.

Listerine use was associated with an increased abundance of common oral opportunistic bacteria previously reported to be enriched in periodontal diseases, oesophageal and colorectal cancer, and systemic diseases. These findings suggest that the regular use of Listerine mouthwash should be carefully considered.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): antimicrobial resistance , Listerine , microbiome and mouthwash
© 2024 The Authors
Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.001830
2024-06-04
2024-09-17
Loading full text...

Full text loading...

References

  1. Caselli E, Fabbri C, D’Accolti M, Soffritti I, Bassi C et al. Defining the oral microbiome by whole-genome sequencing and resistome analysis: the complexity of the healthy picture. BMC Microbiol 2020; 20:1–19 [View Article] [PubMed]
     [Google Scholar]
  2. Dewhirst FE, Chen T, Izard J, Paster BJ, Tanner ACR et al. The human oral microbiome. J Bacteriol 2010; 192:5002–5017 [View Article] [PubMed]
     [Google Scholar]
  3. Aas JA, Paster BJ, Stokes LN, Olsen I, Dewhirst FE. Defining the normal bacterial flora of the oral cavity. J Clin Microbiol 2005; 43:5721–5732 [View Article] [PubMed]
     [Google Scholar]
  4. Saavedra JM, Dattilo A. Microbiota of the intestine: Prebiotics. Encycl Hum Nutr 2012; 3–4:168–174
     [Google Scholar]
  5. Loban GA, Faustova MO, Chereda VV, Ananieva MM. Epidemiological and etiological aspects of dental caries development. Acta fac medic Naissensis 2021; 38:27–34 [View Article]
     [Google Scholar]
  6. Kumar P. Pharmacology of specific drug groups: antibiotic therapy. In Pharmacology and Therapeutics for Dentistry, 7th edn. Elsevier; 2017 pp 457–487
     [Google Scholar]
  7. Mira A, Simon-Soro A, Curtis MA. Role of microbial communities in the pathogenesis of periodontal diseases and caries. J Clin Periodontol 2017; 44:S23–S38 [View Article] [PubMed]
     [Google Scholar]
  8. Kilian M, Chapple ILC, Hannig M, Marsh PD, Meuric V et al. The oral microbiome - an update for oral healthcare professionals. Br Dent J 2016; 221:657–666 [View Article] [PubMed]
     [Google Scholar]
  9. Kapil V, Webb AJ, Ahluwalia A. Inorganic nitrate and the cardiovascular system. Heart 2010; 96:1703–1709 [View Article] [PubMed]
     [Google Scholar]
  10. Rosier BT, Moya-Gonzalvez EM, Corell-Escuin P, Mira A. Isolation and characterization of nitrate-reducing bacteria as potential probiotics for oral and systemic health. Front Microbiol 2020; 11: [View Article] [PubMed]
     [Google Scholar]
  11. Kilian M. The oral microbiome - friend or foe?. Eur J Oral Sci 2018; 126:5–12 [View Article] [PubMed]
     [Google Scholar]
  12. Tartaglia GM, Kumar S, Fornari CD, Corti E, Connelly ST. Mouthwashes in the 21st century: a narrative review about active molecules and effectiveness on the periodontal outcomes. Expert Opin Drug Deliv 2017; 14:973–982 [View Article] [PubMed]
     [Google Scholar]
  13. Brookes ZLS, Bescos R, Belfield LA, Ali K, Roberts A. Current uses of chlorhexidine for management of oral disease: a narrative review. J Dent 2020; 103:103497 [View Article] [PubMed]
     [Google Scholar]
  14. Chow EPF, Williamson DA, Hocking JS, Law MG, Maddaford K et al. Antiseptic mouthwash for gonorrhoea prevention (OMEGA): a randomised, double-blind, parallel-group, multicentre trial. Lancet Infect Dis 2021; 21:647–656 [View Article] [PubMed]
     [Google Scholar]
  15. Van Dijck C, Tsoumanis A, Rotsaert A, Vuylsteke B, Van den Bossche D et al. Antibacterial mouthwash to prevent sexually transmitted infections in men who have sex with men taking HIV pre-exposure prophylaxis (PReGo): a randomised, placebo-controlled, crossover trial. Lancet Infect Dis 2021; 21:657–667 [View Article] [PubMed]
     [Google Scholar]
  16. Zhang L, Regan DG, Chow EPF, Gambhir M, Cornelisse V et al. Neisseria gonorrhoeae transmission among men who have sex with men: an anatomical site-specific mathematical model evaluating the potential preventive impact of mouthwash. Sex Transm Dis 2017; 44:586–592 [View Article] [PubMed]
     [Google Scholar]
  17. Fairley CK, Zhang L, Chow EPF. New thinking on gonorrhoea control in MSM: are antiseptic mouthwashes the answer?. Curr Opin Infect Dis 2018; 31:45–49 [View Article] [PubMed]
     [Google Scholar]
  18. Chow EPF, Maddaford K, Trumpour S, Fairley CK. Translating mouthwash use for gonorrhoea prevention into a public health campaign: identifying current knowledge and research gaps. Sex Health 2019; 16:433–441 [View Article] [PubMed]
     [Google Scholar]
  19. Williams E, Fairley CK, Williamson D. Novel strategies for prevention and treatment of antimicrobial resistance in sexually-transmitted infections. Curr Opin Infect Dis 2021; 34:591–598 [View Article] [PubMed]
     [Google Scholar]
  20. Weinstock H, Workowski KA. Pharyngeal gonorrhea: an important reservoir of infection?. Clin Infect Dis 2009; 49:1798–1800 [View Article] [PubMed]
     [Google Scholar]
  21. Morris SR, Klausner JD, Buchbinder SP, Wheeler SL, Koblin B et al. Prevalence and incidence of pharyngeal gonorrhea in a longitudinal sample of men who have sex with men: the EXPLORE study. Clin Infect Dis 2006; 43:1284–1289 [View Article] [PubMed]
     [Google Scholar]
  22. Unemo M, Shafer WM. Antimicrobial resistance in Neisseria gonorrhoeae in the 21st century: past, evolution, and future. Clin Microbiol Rev 2014; 27:587–613 [View Article] [PubMed]
     [Google Scholar]
  23. Kolokythas A. Mouthwash use in general population: results from adult dental long-term surgical complications in the oral cancer patient: a health survey in Grampian, Scotland. J Oral Maxillofac Res 2010; 1:1–9 [View Article]
     [Google Scholar]
  24. Joshipura KJ, Muñoz-Torres FJ, Morou-Bermudez E, Patel RP. Over-the-counter mouthwash use and risk of pre-diabetes/diabetes. Nitric Oxide - Biol Chem 2017; 71:14–20 [View Article] [PubMed]
     [Google Scholar]
  25. Koopman JE, Van Der Kaaij NCW, Buijs MJ, Elyassi Y, van der Veen MH et al. The effect of fixed orthodontic appliances and fluoride mouthwash on the oral microbiome of adolescents - a randomized controlled clinical trial. PLoS One 2015; 10:1–17 [View Article] [PubMed]
     [Google Scholar]
  26. Plummer EL, Maddaford K, Murray GL, Fairley CK, Pasricha S et al. The impact of mouthwash on the oropharyngeal microbiota of men who have sex with men: a substudy of the OMEGA trial. Microbiol Spectr 2022; 10:e0175721 [View Article] [PubMed]
     [Google Scholar]
  27. Bescos R, Ashworth A, Cutler C, Brookes ZL, Belfield L et al. Effects of Chlorhexidine mouthwash on the oral microbiome. Sci Rep 2020; 10:1–8 [View Article] [PubMed]
     [Google Scholar]
  28. Tribble GD, Angelov N, Weltman R, Wang BY, Eswaran SV et al. Frequency of tongue cleaning impacts the human tongue microbiome composition and enterosalivary circulation of nitrate. Front Cell Infect Microbiol 2019; 9:1–16 [View Article] [PubMed]
     [Google Scholar]
  29. Laumen JGE, Van Dijck C, Abdellati S, De Baetselier I, Serrano G et al. Antimicrobial susceptibility of commensal Neisseria in a general population and men who have sex with men in Belgium. Sci Rep 2022; 12:1–10 [View Article] [PubMed]
     [Google Scholar]
  30. Bolger AM, Lohse M, Usadel B. Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 2014; 30:2114–2120 [View Article] [PubMed]
     [Google Scholar]
  31. Li H, Handsaker B, Wysoker A, Fennell T, Ruan J et al. The sequence alignment/map format and SAMtools. Bioinformatics 2009; 25:2078–2079 [View Article] [PubMed]
     [Google Scholar]
  32. Li H, Durbin R. Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics 2009; 25:1754–1760 [View Article] [PubMed]
     [Google Scholar]
  33. Wood DE, Lu J, Langmead B. Improved metagenomic analysis with Kraken 2. Genome Biol 2019; 20:1–13 [View Article] [PubMed]
     [Google Scholar]
  34. Lu J, Breitwieser FP, Thielen P, Salzberg SL. Bracken: estimating species abundance in metagenomics data. PeerJ Comput Sci 2017; 3:1–17 [View Article]
     [Google Scholar]
  35. Gloor GB, Macklaim JM, Pawlowsky-Glahn V, Egozcue JJ. Microbiome datasets are compositional: and this is not optional. Front Microbiol 2017; 8:1–6 [View Article] [PubMed]
     [Google Scholar]
  36. Wickham H. ggplot2. WIREs Computational Stats 2011; 3:180–185 [View Article]
     [Google Scholar]
  37. Gu Z, Eils R, Schlesner M. Complex heatmaps reveal patterns and correlations in multidimensional genomic data. Bioinformatics 2016; 32:2847–2849 [View Article] [PubMed]
     [Google Scholar]
  38. Lahti L, Shetty S. Tools for microbiome analysis in R; 2017 http://microbiome.github.com/microbiome
  39. McMurdie PJ, Holmes S. Phyloseq: an R package for reproducible interactive analysis and graphics of microbiome census data. PLoS One 2013; 8:e61217 [View Article] [PubMed]
     [Google Scholar]
  40. Xu S, Li Z, Tang W, Dai Z, Zhou L et al. MicrobiotaProcess: a comprehensive R package for managing and analyzing microbiome and other ecological data within the tidy framework. Res Sq 2022 [View Article]
     [Google Scholar]
  41. Jari Oksanen FGB, Michael Friendly R, Kindt PL, McGlinn D, Peter R. Minchin RB et al. Vegan: Community Ecology Package Community Ecology Package; 2020
     [Google Scholar]
  42. Greg Gloor Aldex2: ANOVA-like differential expression tool for compositional data; 2017
  43. Mandal S, Van Treuren W, White RA, Eggesbø M, Knight R et al. Analysis of composition of microbiomes: a novel method for studying microbial composition. Microb Ecol Health Dis 2015; 26:27663 [View Article] [PubMed]
     [Google Scholar]
  44. Salter SJ, Cox MJ, Turek EM, Calus ST, Cookson WO et al. Reagent and laboratory contamination can critically impact sequence-based microbiome analyses. BMC Biol 2014; 12:1–12 [View Article] [PubMed]
     [Google Scholar]
  45. Amaral CDSF, Da Silva-Boghossian CMI, Leão ATT, Colombo APV. Evaluation of the subgingival microbiota of alcoholic and non-alcoholic individuals. J Dent 2011; 39:729–738 [View Article] [PubMed]
     [Google Scholar]
  46. Merchant AT, Park YM, Dodhia S, Shrestha D, Choi YH et al. Cross-sectional analysis of alcohol intake and serum antibodies to oral microorganisms. JDR Clin Transl Res 2017; 2:168–178 [View Article]
     [Google Scholar]
  47. Morita E, Narikiyo M, Yokoyama A, Yano A, Kamoi K et al. Predominant presence of Streptococcus anginosus in the saliva of alcoholics. Oral Microbiol Immunol 2005; 20:362–365 [View Article] [PubMed]
     [Google Scholar]
  48. Barb JJ, Maki KA, Kazmi N, Meeks BK, Krumlauf M et al. The oral microbiome in alcohol use disorder: a longitudinal analysis during inpatient treatment. J Oral Microbiol 2022; 14:2004790 [View Article] [PubMed]
     [Google Scholar]
  49. Jacobs JA, Schot CS, Schouls LM. The Streptococcus anginosus species comprises five 16S rRNA ribogroups with different phenotypic characteristics and clinical relevance. Int J Syst Evol Microbiol 2000; 50:1073–1079 [View Article] [PubMed]
     [Google Scholar]
  50. McIlvanna E, Linden GJ, Craig SG, Lundy FT, James JA. Fusobacterium nucleatum and oral cancer: a critical review. BMC Cancer 2021; 21:1–11 [View Article] [PubMed]
     [Google Scholar]
  51. Narikiyo M, Tanabe C, Yamada Y, Igaki H, Tachimori Y et al. Frequent and preferential infection of Treponema denticola, Streptococcus mitis, and Streptococcus anginosus in esophageal cancers. Cancer Sci 2004; 95:569–574 [View Article] [PubMed]
     [Google Scholar]
  52. Tateda M, Shiga K, Saijo S, Sone M, Hori T et al. Streptococcus anginosus in head and neck squamous cell carcinoma: implication in carcinogenesis. Int J Mol Med 2000; 6:699–703 [View Article] [PubMed]
     [Google Scholar]
  53. Jackson DS, Welch DF, Pickett DA, Mukwaya GM, Kuhls TL. Suppurative infections in children caused by non-beta-hemolytic members of the Streptococcus milleri group. Pediatr Infect Dis J 1995; 14:80–82 [View Article] [PubMed]
     [Google Scholar]
  54. Belko J, Goldmann DA, Macone A, Zaidi AKM. Clinically significant infections with organisms of the Streptococcus milleri group. Pediatr Infect Dis J 2002; 21:715–723 [View Article] [PubMed]
     [Google Scholar]
  55. Doern CD, Burnham CAD. It’s not easy being green: the viridans group streptococci, with a focus on pediatric clinical manifestations. J Clin Microbiol 2010; 48:3829–3835 [View Article] [PubMed]
     [Google Scholar]
  56. Nagashima H, Takao A, Maeda N. Abscess forming ability of streptococcus milleri group: synergistic effect with Fusobacterium nucleatum. Microbiol Immunol 1999; 43:207–216 [View Article] [PubMed]
     [Google Scholar]
  57. Mukae H, Noguchi S, Naito K, Kawanami T, Yamasaki K et al. The importance of obligate anaerobes and the Streptococcus anginosus group in pulmonary abscess: a clone library analysis using Bronchoalveolar lavage fluid. Respiration 2016; 92:80–89 [View Article] [PubMed]
     [Google Scholar]
  58. Brennan CA, Garrett WS. Fusobacterium nucleatum - symbiont, opportunist and oncobacterium. Nat Rev Microbiol 2019; 17:156–166 [View Article] [PubMed]
     [Google Scholar]
  59. Wang S, Liu Y, Li J, Zhao L, Yan W et al. Fusobacterium nucleatum acts as a pro-carcinogenic bacterium in colorectal cancer: from association to causality. Front Cell Dev Biol 2021; 9:1–15 [View Article] [PubMed]
     [Google Scholar]
  60. Hsueh CY, Huang Q, Gong H, Shen Y, Sun J et al. A positive feed-forward loop between Fusobacterium nucleatum and ethanol metabolism reprogramming drives laryngeal cancer progression and metastasis. iScience 2022; 25:103829 [View Article] [PubMed]
     [Google Scholar]
  61. Paolo Boffetta MH. Alcohol and Cancer. The Lancet 1976; 307:1243 [View Article]
     [Google Scholar]
  62. Wight AJ, Ogden GR. Possible mechanisms by which alcohol may influence the development of oral cancer-a review. Oral Oncol 1998; 34:441–447 [View Article] [PubMed]
     [Google Scholar]
  63. Kurkivuori J, Salaspuro V, Kaihovaara P, Kari K, Rautemaa R et al. Acetaldehyde production from ethanol by oral streptococci. Oral Oncol 2007; 43:181–186 [View Article] [PubMed]
     [Google Scholar]
  64. Homann N, Jousimies-Somer H, Jokelainen K, Heine R, Salaspuro M. High acetaldehyde levels in saliva after ethanol consumption: methodological aspects and pathogenetic implications. Carcinogenesis 1997; 18:1739–1743 [View Article] [PubMed]
     [Google Scholar]
  65. Ustrell-Borràs M, Traboulsi-Garet B, Gay-Escoda C. Alcohol-based mouthwash as a risk factor of oral cancer: a systematic review. Med Oral Patol Oral Cir Bucal 2020; 25:e1–e12 [View Article] [PubMed]
     [Google Scholar]
  66. Eliot MN, Michaud DS, Langevin SM, McClean MD, Kelsey KT. Periodontal disease and mouthwash use are risk factors for head and neck squamous cell carcinoma. Cancer Causes Control 2013; 24:1315–1322 [View Article] [PubMed]
     [Google Scholar]
  67. Doel JJ, Benjamin N, Hector MP, Rogers M, Allaker RP. Evaluation of bacterial nitrate reduction in the human oral cavity. Eur J Oral Sci 2005; 113:14–19 [View Article] [PubMed]
     [Google Scholar]
  68. Hyde ER, Andrade F, Vaksman Z, Parthasarathy K, Jiang H et al. Metagenomic analysis of nitrate-reducing bacteria in the oral cavity: implications for nitric oxide homeostasis. PLoS One 2014; 9:e88645 [View Article] [PubMed]
     [Google Scholar]
  69. Bryan NS, Tribble G, Angelov N. Oral microbiome and nitric oxide: the missing link in the management of blood pressure. Curr Hypertens Rep 2017; 19:33 [View Article] [PubMed]
     [Google Scholar]
  70. Hezel MP, Weitzberg E. The oral microbiome and nitric oxide homoeostasis. Oral Dis 2015; 21:7–16 [View Article] [PubMed]
     [Google Scholar]
  71. Govoni M, Jansson EA, Weitzberg E, Lundberg JO. The increase in plasma nitrite after a dietary nitrate load is markedly attenuated by an antibacterial mouthwash. Nitric Oxide - Biol Chem 2008; 19:333–337 [View Article] [PubMed]
     [Google Scholar]
  72. Woessner M, Smoliga JM, Tarzia B, Stabler T, Van Bruggen M et al. A stepwise reduction in plasma and salivary nitrite with increasing strengths of mouthwash following a dietary nitrate load. Nitric Oxide Biol Chem 2016; 54:1–7 [View Article] [PubMed]
     [Google Scholar]
  73. Kapil V, Haydar SMA, Pearl V, Lundberg JO, Weitzberg E et al. Physiological role for nitrate-reducing oral bacteria in blood pressure control. Free Radic Biol Med 2013; 55:93–100 [View Article] [PubMed]
     [Google Scholar]
  74. Mitsui T, Harasawa R. The effects of essential oil, povidone-iodine, and chlorhexidine mouthwash on salivary nitrate/nitrite and nitrate-reducing bacteria. J Oral Sci 2017; 59:597–601 [View Article] [PubMed]
     [Google Scholar]
  75. Alzahrani HS, Jackson KG, Hobbs DA, Lovegrove JA. The role of dietary nitrate and the oral microbiome on blood pressure and vascular tone. Nutr Res Rev 2021; 34:222–239 [View Article] [PubMed]
     [Google Scholar]
  76. Brookes ZLS, Belfield LA, Ashworth A, Casas-Agustench P, Raja M et al. Effects of chlorhexidine mouthwash on the oral microbiome. J Dent 2021; 113:103768 [View Article] [PubMed]
     [Google Scholar]
  77. Al-Kamel A, Baraniya D, Al-Hajj WA, Halboub E, Abdulrab S et al. Subgingival microbiome of experimental gingivitis: shifts associated with the use of chlorhexidine and N-acetyl cysteine mouthwashes. J Oral Microbiol 2019; 11:1608141 [View Article] [PubMed]
     [Google Scholar]
  78. Exterkate RAM, Zaura E, Brandt BW, Buijs MJ, Koopman JE et al. The effect of propidium monoazide treatment on the measured bacterial composition of clinical samples after the use of a mouthwash. Clin Oral Investig 2015; 19:813–822 [View Article] [PubMed]
     [Google Scholar]
  79. Durazzi F, Sala C, Castellani G, Manfreda G, Remondini D et al. Comparison between 16S rRNA and shotgun sequencing data for the taxonomic characterization of the gut microbiota. Sci Rep 2021; 11:1–10 [View Article] [PubMed]
     [Google Scholar]
  80. Van Dijck C, Kenyon C, Cuylaerts V, Sollie P, Spychala A et al. The development of mouthwashes without anti-gonococcal activity for controlled clinical trials: an in vitro study. F1000Res 2020; 8:1–11 [View Article] [PubMed]
     [Google Scholar]
  81. Xu X, He J, Xue J, Wang Y, Li K et al. Oral cavity contains distinct niches with dynamic microbial communities. Environ Microbiol 2015; 17:699–710 [View Article] [PubMed]
     [Google Scholar]
/content/journal/jmm/10.1099/jmm.0.001830
Loading
The effect of daily usage of Listerine Cool Mint mouthwash on the oropharyngeal microbiome: a substudy of the PReGo trial
J. Med. Microbiol. 73, 001830 (2024); https://doi.org/10.1099/jmm.0.001830
/content/journal/jmm/10.1099/jmm.0.001830
/content/journal/jmm/10.1099/jmm.0.001830
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited Most Cited RSS feed

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