Colonization, safety, and tolerability study of the Streptococcus salivarius 24SMBc nasal spray for its application in upper respiratory tract infections

  • M. SantagatiEmail author
  • M. Scillato
  • N. Muscaridola
  • V. Metoldo
  • I. La Mantia
  • S. Stefani
Original Article


Streptococcus salivarius, a non-pathogenic species and the predominant colonizer of the oral microbiota, finds a wide application in the prevention of upper respiratory tract infections, also reducing the frequency of their main pathogens. In this pilot study, the primary objective was to evaluate the safety and tolerability of a nasal spray, S. salivarius 24SMBc, as a medical device in a clinical study involving 20 healthy adult subjects. The secondary aim was to determine the ability of colonization assessed by molecular fingerprinting. Twenty healthy adult subjects, aged between 30 and 54 years, without a medical history of recurrent otitis media, were enrolled. All patient characteristics fulfilled the inclusion criteria. All subjects were treated daily for 3 days with the nasal spray containing S. salivarius 24SMBc at a concentration of 5 × 109 colony-forming units (CFU)/ml. The persistence of S. salivarius in the nasopharynx was investigated by the antagonism test and random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR). The tolerability and safety were clinically assessed by clinical examinations during treatment. Our results demonstrate the capability of S. salivarius 24SMBc to colonize the rhinopharynx tissues in 95 % of subjects and persist in 55 % of them after 6 days from the last dose of the formulation, maintaining a concentration of 105 CFU/ml. The treatment was well tolerated by all healthy patients and no adverse effects were found. The topical application of streptococcal probiotics is a relatively undeveloped field but is becoming an attractive approach for both prevention and therapy, especially for pediatric age patients. S. salivarius 24SMBc possess characteristics making this strain suitable for use in bacteriotherapy.


Nasal Spray Moraxella Catarrhalis Oral Microbiota Recurrent Otitis Medium Molecular Fingerprinting 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This study was supported by a project funded by the Italian Ministry of Education, University and Research (MIUR), PON_02589 and a research grant from DMG Italia. The nasal spray as a medical device was kindly provided by DMG Italia. We thank the Scientific Bureau of the University of Catania for the language support.

Conflict of interest

The authors declare that there is no conflict of interest.


  1. 1.
    Lenoir-Wijnkoop I, Sanders ME, Cabana MD, Caglar E, Corthier G, Rayes N, Sherman PM, Timmerman HM, Vaneechoutte M, Van Loo J, Wolvers DA (2007) Probiotic and prebiotic influence beyond the intestinal tract. Nutr Rev 11(65):469–489CrossRefGoogle Scholar
  2. 2.
    Gareau MG, Sherman PM, Walker WA (2010) Probiotics and the gut microbiota in intestinal health and disease. Nat Rev Gastroenterol Hepatol 7(9):503–514, reviewCrossRefPubMedGoogle Scholar
  3. 3.
    Ashraf R, Shah NP (2014) Immune system stimulation by probiotic microorganisms. Crit Rev Food Sci Nutr 54:938–956CrossRefPubMedGoogle Scholar
  4. 4.
    Koboziev I, Reinoso Webb C, Furr KL, Grisham MB (2014) Role of the enteric microbiota in intestinal homeostasis and inflammation. Free Radic Biol Med 68:122–133CrossRefPubMedGoogle Scholar
  5. 5.
    Liu S, Hu P, Du X, Zhou T, Pei X (2013) Lactobacillus rhamnosus GG supplementation for preventing respiratory infections in children: a meta-analysis of randomized, placebo-controlled trials. Indian Pediatr 50(4):377–381CrossRefPubMedGoogle Scholar
  6. 6.
    Cohen R, Martin E, de La Rocque F, Thollot F, Pecquet S, Werner A, Boucherat M, Varon E, Bingen E, Levy C (2013) Probiotics and prebiotics in preventing episodes of acute otitis media in high-risk children: a randomized, double-blind, placebo-controlled study. Pediatr Infect Dis J 32(8):810–814PubMedGoogle Scholar
  7. 7.
    Hatakka K, Blomgren K, Pohjavuori S, Kaijalainen T, Poussa T, Leinonen M, Korpela R, Pitkäranta A (2007) Treatment of acute otitis media with probiotics in otitis-prone children a double-blind, placebo-controlled randomised study. Clin Nutr 26(3):314–321Google Scholar
  8. 8.
    Wescombe PA, Hale JD, Heng NC, Tagg JR (2012) Developing oral probiotics from Streptococcus salivarius. Future Microbiol 7:1355–1371CrossRefPubMedGoogle Scholar
  9. 9.
    Monasta L, Ronfani L, Marchetti F, Montico M, Vecchi Brumatti L, Bavcar A, Grasso D, Barbiero C, Tamburlini G (2012) Burden of disease caused by otitis media: systematic review and global estimates. PLoS One 7:e36226CrossRefPubMedCentralPubMedGoogle Scholar
  10. 10.
    Lieberthal AS, Carroll AE, Chonmaitree T, Ganiats TG, Hoberman A, Jackson MA, Joffe MD, Miller DT, Rosenfeld RM, Sevilla XD, Schwartz RH, Thomas PA, Tunkel DE; American Academy of Pediatrics Subcommittee on Management of Acute Otitis Media (2013) The diagnosis and management of acute otitis media. Pediatrics 131:e964–e999CrossRefPubMedGoogle Scholar
  11. 11.
    Marchisio P, Claut L, Rognoni A, Esposito S, Passali D, Bellussi L, Drago L, Pozzi G, Mannelli S, Schito G, Principi N (2003) Differences in nasopharyngeal bacterial flora in children with nonsevere recurrent acute otitis media and chronic otitis media with effusion: implications for management. Pediatr Infect Dis J 22:262–268PubMedGoogle Scholar
  12. 12.
    John M, Dunne EM, Licciardi PV, Satzke C, Wijburg O, Robins-Browne RM, O’Leary S (2013) Otitis media among high-risk populations: can probiotics inhibit Streptococcus pneumoniae colonisation and the risk of disease? Eur J Clin Microbiol Infect Dis 32(9):1101–1110, reviewCrossRefPubMedGoogle Scholar
  13. 13.
    Tagg JR (2004) Prevention of streptococcal pharyngitis by anti-Streptococcus pyogenes bacteriocin-like inhibitory substances (BLIS) produced by Streptococcus salivarius. Indian J Med Res 119(Suppl):13–16PubMedGoogle Scholar
  14. 14.
    Burton JP, Chilcott CN, Moore CJ, Speiser G, Tagg JR (2006) A preliminary study of the effect of probiotic Streptococcus salivarius K12 on oral malodour parameters. J Appl Microbiol 100(4):754–764CrossRefPubMedGoogle Scholar
  15. 15.
    Santagati M, Scillato M, Patanè F, Aiello C, Stefani S (2012) Bacteriocin-producing oral streptococci and inhibition of respiratory pathogens. FEMS Immunol Med Microbiol 65(1):23–31CrossRefPubMedGoogle Scholar
  16. 16.
    Stol K, Verhaegh SJ, Graamans K, Engel JA, Sturm PD, Melchers WJ, Meis JF, Warris A, Hays JP, Hermans PW (2013) Microbial profiling does not differentiate between childhood recurrent acute otitis media and chronic otitis media with effusion. Int J Pediatr Otorhinolaryngol 77(4):488–493CrossRefPubMedGoogle Scholar
  17. 17.
    Célind J, Södermark L, Hjalmarson O (2014) Adherence to treatment guidelines for acute otitis media in children. The necessity of an effective strategy of guideline implementation. Int J Pediatr Otorhinolaryngol 78(7):1128–1132CrossRefPubMedGoogle Scholar
  18. 18.
    Tagg JR, Bannister LV (1979) “Fingerprinting” beta-haemolytic streptococci by their production of and sensitivity to bacteriocine-like inhibitors. J Med Microbiol 12:397–411CrossRefPubMedGoogle Scholar
  19. 19.
    Truong TL, Ménard C, Mouton C, Trahan L (2000) Identification of mutans and other oral streptococci by random amplified polymorphic DNA analysis. J Med Microbiol 49:63–71CrossRefPubMedGoogle Scholar
  20. 20.
    Conticello S, Saita V, La Mantia I, Ferlito S (1989) Endoscopy of the eustachian tube: use of the fiberscope and the telescope. Arch Otorhinolaryngol 246:256–258, Sprinter-Verlag EdCrossRefPubMedGoogle Scholar
  21. 21.
    Serra A, Grillo C, La Mantia I, Cipri R, Vancheri M, Saita V (1992) Naso-pharyngo-laryngoscopie et rhinomanométrie avec endoscope flexible dans l’étude de l’hypertrophie adénoïde. Acta Endoscopica Belg 22:381–384CrossRefGoogle Scholar
  22. 22.
    Human Microbiome Project Consortium (2012) Structure, function and diversity of the healthy human microbiome. Nature 486:207–214CrossRefGoogle Scholar
  23. 23.
    Aas JA, Paster BJ, Stokes LN, Olsen I, Dewhirst FE (2005) Defining the normal bacterial flora of the oral cavity. J Clin Microbiol 43(11):5721–5732CrossRefPubMedCentralPubMedGoogle Scholar
  24. 24.
    Soccol CR, Porto de Souza Vandenberghe L, Spier MR, Pedroni Medeiros AB, Yamaguishi CT, De Dea Lindner J, Pandey A, Thomaz-Soccol V (2010) The potential of probiotics: a review. Food Technol Biotech 48(4):413–434Google Scholar
  25. 25.
    Caglar E, Kargul B, Tanboga I (2005) Bacteriotherapy and probiotics’ role on oral health. Oral Dis 11(3):131–137CrossRefPubMedGoogle Scholar
  26. 26.
    Di Pierro F, Colombo M, Zanvit A, Risso P, Rottoli AS (2014) Use of Streptococcus salivarius K12 in the prevention of streptococcal and viral pharyngotonsillitis in children. Drug Healthc Patient Saf 6:15–20CrossRefPubMedCentralPubMedGoogle Scholar
  27. 27.
    Walls T, Power D, Tagg J (2003) Bacteriocin-like inhibitory substance (BLIS) production by the normal flora of the nasopharynx: potential to protect against otitis media? J Med Microbiol 52(Pt 9):829–833CrossRefPubMedGoogle Scholar
  28. 28.
    Roos K, Håkansson EG, Holm S (2001) Effect of recolonisation with “interfering” alpha streptococci on recurrences of acute and secretory otitis media in children: randomised placebo controlled trial. BMJ 322(7280):210–212CrossRefPubMedCentralPubMedGoogle Scholar
  29. 29.
    Tano K, Grahn Håkansson E, Holm SE, Hellström S (2002) A nasal spray with alpha-haemolytic streptococci as long term prophylaxis against recurrent otitis media. Int J Pediatr Otorhinolaryngol 62(1):17–23CrossRefPubMedGoogle Scholar
  30. 30.
    Skovbjerg S, Roos K, Holm SE, Grahn Håkansson E, Nowrouzian F, Ivarsson M, Adlerberth I, Wold AE (2009) Spray bacteriotherapy decreases middle ear fluid in children with secretory otitis media. Arch Dis Child 94(2):92–98CrossRefPubMedGoogle Scholar
  31. 31.
    Gao Z, Kang Y, Yu J, Ren L (2014) Human pharyngeal microbiome may play a protective role in respiratory tract infections. Genomics Proteomics Bioinformatics 12(3):144–150CrossRefPubMedCentralPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • M. Santagati
    • 1
    Email author
  • M. Scillato
    • 1
  • N. Muscaridola
    • 1
  • V. Metoldo
    • 1
  • I. La Mantia
    • 2
  • S. Stefani
    • 1
  1. 1.LabMMAR, Department of Biomedical and Biotechnological Sciences, Section of MicrobiologyUniversity of CataniaCataniaItaly
  2. 2.Department of Medical Sciences, Surgical and Advanced Technologies, GF IngrassiaUniversity of CataniaCataniaItaly

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