Abstract
The aim of this study was to determine the prevalence of Streptococcus mutans and its serotypes in samples from oral cavity of young Galician population and their relationship with the oral health state. The variables generally associated with dental caries, such as salivary flow rate, buffering capacity, eating habits, and lifestyle, were also analysed. No relationship was found between the variables studied and the presence of S. mutans in the oral cavity or the existence of dental caries. Presumptive strains of S. mutans were isolated from saliva samples from 48% of the analysed population. The use of conventional microbiological methods, API 20 Strep system, and species-specific polymerase chain reaction (PCR) allowed to substantiate the identity of the strains as S. mutans. Multiplex PCR protocols, developed in this study for the simultaneous detection of S. mutans and serotypes c, e, and f and for detection of S. mutans and serotype k, also confirmed this result and demonstrated that serotype c was predominant in the studied young Galician population (86%). Serotypes e (8%), k (3%), and f (2%) were also detected. Serotype c was detected in carious and caries-free subjects, while the remaining serotypes were only found in subjects with caries.
Similar content being viewed by others
Data availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on request.
References
Abelson DC (1989) The effect of hydrogenated starch hydrolysates on plaque pH in vivo. Clin Prev Dent 11:20–23
Al-mudallal NHA, Al-jumaily EFA, Al-shaibany AA, Engineering G, Studies P (2008) Isolation and identification of mutan’s streptococci. J Al-Nahrain Univ 11:98–105
Banas JA (2004) Virulence properties of Streptococcus mutans. Front Biosci 9(1267):1277. https://doi.org/10.2741/1305
Carletto-Körber FP, González-Ittig RE, Jimenez MG, Cornejo LS (2015) Serotype diversity of Streptococcus mutans and caries activity in children in Argentina. Eur J Paediatr Dent 16:177–180
Childers NK, Osgood RC, Hsu KL, Manmontri C, Momeni SS, Mahtani H, Cutter GR, Ruby JD (2011) Real-time quantitative polymerase chain reaction for enumeration of Streptococcus mutans from oral samples. PLoS One 119:447–454. https://doi.org/10.1371/journal.pone.0178059
Clinical Laboratory Standards Institute (2008) Interpretive criteria for identification of bacteria and fungi by DNA target sequencing: approved guideline MM18-a, 2nd edn. Wayne, USA
Cunha-Cruz J, Scott J, Rothen M, Mancl L, Lawhorn T, Brossel K, Berg J (2013) Salivary characteristics and dental caries: evidence from general dental practices. J Am Dent Assoc 144:31–40. https://doi.org/10.1038/jid.2014.371
De La Higuera A, Gutiérrez J, Liébana J, Garcia-Mendoza A, Castillo A (1999) A new biotyping method for Streptococcus mutans with the API ZYM system. Clin Microbiol Infect 5:88–91
Dodds MWJ, Johnson DA, Yeh CK (2005) Health benefits of saliva: a review. J Dent 33:223–233. https://doi.org/10.1016/j.jdent.2004.10.009
Fontana C, Favaro M, Pelliccioni M, Pistoia ES, Favalli C (2005) Use of the MicroSeq 500 16S rRNA gene-based sequencing for identification of bacterial isolates that commercial automated systems failed to identify correctly. J Clin Microbiol 43:615–619. https://doi.org/10.1128/JCM.43.2.615-619.2005
Garnier F, Gerbaud G, Courvalin P, Galimand M (1997) Identification of clinically relevant viridans group streptococci to the species level by PCR. J Clin Microbiol 35:2337–2341
Hirasawa M, Takada K (2003) A new selective medium for Streptococcus mutans and the distribution of S. mutans and S. sobrinus and their serotypes in dental plaque. Caries Res 37:212–217. https://doi.org/10.1159/000070447
Hoshino T, Kawaguchi M, Shimizu N, Hoshino N, Ooshima T, Fujiwara T (2004) PCR detection and identification of oral streptococci in saliva samples using gtf genes. Diagn Microbiol Infect Dis 48:195–199. https://doi.org/10.1016/j.diagmicrobio.2003.10.002
Hung WC, Tsai JC, Hsueh PR, Chia JS, Teng LJ (2005) Species identification of mutans streptococci by groESL gene sequence. J Med Microbiol 54:857–862. https://doi.org/10.1099/jmm.0.46180-0
Krzyściak W, Kościelniak D, Papiez M, Jurczak A, Vyhouskaya P (2017) Methods of biotyping of Streptococcus mutans species with the routine test as a prognostic value in early childhood caries. Evid Based Complement Alternat Med 2017:1–15. https://doi.org/10.1155/2017/6859543
MacFaddin JF (2003) Pruebas bioquímicas para la identificación de bacterias de importancia clínica, 3rd edn. Ed. Medica Panamericana, Buenos Aires
Meurman JH, Rantonen P (1994) Salivary flow-rate, buffering capacity, and yeast counts in 187 consecutive adult patients from Kuopio, Finland. Scand J Dent Res 102:229–234
Momeni SS, Whiddon J, Cheon K, Moser SA, Childers NK (2015) Assessment of clonality and serotypes of Streptococcus mutans among children by multilocus sequence typing. Eur J Oral Sci 123:416–424. https://doi.org/10.1111/eos.12221
Momeni SS, Ghazal T, Grenett H, Whiddon J, Moser SA, Childers NK (2019) Streptococcus mutans serotypes and collagen-binding proteins Cnm/Cbm in children with caries analysed by PCR. Mol Oral Microbiol 34:64–73. https://doi.org/10.1111/omi.12254
Nakano K, Ooshima T (2009) Serotype classification of Streptococcus mutans and its detection outside the oral cavity. Future Microbiol 4:891–902. https://doi.org/10.2217/fmb.09.64
Nakano K, Nomura R, Shimizu N, Nakagawa I, Hamada S, Ooshima T (2004) Development of a PCR method for rapid identification of new Streptococcus mutans serotype k strains. J Clin Microbiol 42:4925–4930. https://doi.org/10.1128/JCM.42.11.4925-4930.2004
Nakano K, Nemoto H, Nomura R, Homma H, Yoshioka H, Shudo Y, Hata H, Toda K, Taniguchi K, Amano A, Ooshima T (2007) Serotype distribution of Streptococcus mutans a pathogen of dental caries in cardiovascular specimens from Japanese patients. J Med Microbiol 56:551–556. https://doi.org/10.1099/jmm.0.47051-0
Nakano K, Nomura R, Matsumoto M, Ooshima T (2010) Roles of oral bacteria in cardiovascular diseases - from molecular mechanisms to clinical cases: cell-surface structures of novel serotype k Streptococcus mutans strains and their correlation to virulence. J Pharmacol Sci 113:120–125. https://doi.org/10.1254/jphs.09R24FM
Nakano K, Nakagawa I, Alaluusua S, Ooshima T (2013) Molecular typing of Streptococcus mutans. In: de Filippis I, Mckee M (eds) Molecular typing in bacterial infections, 2nd edn. Humana Press, New York, pp 127–147
Nomura R, Nakano K, Taniguchi N, Lapirattanakul J, Nemoto H, Grönroos L, Alaluusua S, Ooshima T (2009) Molecular and clinical analyses of the gene encoding the collagen-binding adhesin of Streptococcus mutans. J Med Microbiol 58:469–475. https://doi.org/10.1099/jmm.0.007559-0
Oho T, Yamashita Y, Shimazaki Y, Kushiyama M, Koga T (2000) Simple and rapid detection of Streptococcus mutans and Streptococcus sobrinus in human saliva by polymerase chain reaction. Oral Microbiol Immunol 15:258–262. https://doi.org/10.1034/j.1399-302X.2000.150408
Oong EM, Griffin SO, Kohn WG, Gooch BF, Caufield PW (2008) The effect of dental sealants on bacteria levels in caries lesions: a review of the evidence. J Am Dent Assoc 139:271–278. https://doi.org/10.14219/jada.archive.2008.0156
Rao A, Austin R (2014) Serotype specific polymerase chain reaction identifies a higher prevalence of Streptococcus mutans serotype k and e in a random group of children with dental caries from the southern region of India. Contemp Clin Dent 5:296–301. https://doi.org/10.4103/0976-237x.137905
Sánchez AM, Montiel JM, Dasí FF, Almerich SJM (2013) Streptococcus mutans and Streptococcus sobrinus detection by polymerase chain reaction and their relation to dental caries in 12 and 15 year-old schoolchildren in Valencia (Spain). Med Oral Pat Oral y Cir Bucal 18:0–6. https://doi.org/10.4317/medoral.18941
Seki M, Yamashita Y, Shibata Y, Torigoe H, Tsuda H, Maeno M (2006) Effect of mixed mutans streptococci colonization on caries development. Oral Microbiol Immunol 21:47–52. https://doi.org/10.1111/j.1399-302X.2005.00253.x
Shibata Y, Ozaki K, Seki M, Kawato T, Tanaka H, Nakano Y, Yamashita Y (2003) Analysis of loci required for determination of serotype antigenicity in Streptococcus mutans and its clinical utilization. J Clin Microbiol 41:4107–4112. https://doi.org/10.1128/JCM.41.9.4107-4112.2003
Srinivasan R, Karaoz U, Volegova M, MacKichan J, Kato-Maeda M, Miller S, Nadarajan R, Brodie EL, Lynch SV (2015) Use of 16S rRNA gene for identification of a broad range of clinically relevant bacterial pathogens. PLoS One 10:e0117617. https://doi.org/10.1371/journal.pone.0117617
Swedan SF, Obeidat HM, Shakhatreh MAK (2018) Molecular typing and detection of collagen binding genes among Streptococcus mutans isolated from diabetic and non- diabetic individuals in Northern Jordan. Jordan J Biol Sci 11:293–300 http://jjbs.hu.edu.jo/files/v11n3/Paper%20Number%209.pdf
Tenovuo J (1997) Salivary parameters of relevance for assessing caries activity in individuals and populations. Community Dent Oral Epidemiol 25:82–86. https://doi.org/10.1111/j.1600-0528.1997.tb00903
Vos P, Garrity G, Jones D, Krieg N, Ludwig W, Rainey F, Schleifer K, Whitman W (2011) Bergey’s manual of systematic bacteriology: Volume 3: The Firmicutes. Springer, New York
Woo PCY, Teng JLL, Yeung JMY, Tse H, Lau SKP, Yuen KY (2011) Automated identification of medically important bacteria by 16S rRNA gene sequencing using a novel comprehensive database, 16SpathDB. J Clin Microbiol 49:1799–1809. https://doi.org/10.1128/JCM.02350-10
Xu X, Zhou XD, Wu CD (2011) The tea catechin epigallocatechin gallate suppresses cariogenic virulence factors of Streptococcus mutans. Antimicrob Agents Chemother 55:1229–1236. https://doi.org/10.1128/AAC.01016-10
Yoshida A, Suzuki N, Nakano Y, Kawada M, Oho T, Koga T (2003) Development of a 5′ nuclease-based real-time PCR assay for quantitative detection of cariogenic dental pathogens Streptococcus mutans and Streptococcus sobrinus. J Clin Microbiol 41:4438–4441. https://doi.org/10.1128/JCM.41.9.4438-4441.2003
Acknowledgements
Nancy Saltos Rosero thanks to the Secretaría de Educación Superior, Ciencia, Tecnología e Innovación (SENESCYT) of Ecuador for the pre-doctoral scholarship and to Dr. Yoshihisa Yamashita from the Kyushu University of Japan for kindly providing us with S. mutans strains.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Informed consent
All volunteers signed a written informed consent approved by the Bioethics Committee of the Universidade de Santiago de Compostela. Each questionnaire and saliva sample obtained in this study was identified by a number, so that it could not be associated with any person.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Saltos Rosero, N., Seoane Prado, R., Aguilera Guirao, A. et al. Molecular and serological typing of Streptococcus mutans strains isolated from young Galician population: relationship with the oral health status. Int Microbiol 23, 589–596 (2020). https://doi.org/10.1007/s10123-020-00132-2
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10123-020-00132-2