The Antimicrobial Effects of Lactobacillus Plantarum and Lactobacillus Rhamnosus Isolated from Local Kefir Buttermilk on Oral Pathogens

Document Type : Original Article(s)

Authors

1 MSc of Medical Microbiology, Department of Bacteriology and Virology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

2 Assistant Professor, Department of Bacteriology and Virology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

3 PhD Candidate of Medical Bacteriology, Department of Bacteriology and Virology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, Iran

Abstract

Background: Tooth decay is one of the most common diseases, and Streptococcus mutans and Porphyromonas gingivalis have been found to play an effective role in causing advanced tooth decay. With the growing antibiotic resistance of bacteria, the use of probiotics has been investigated to reduce pathogenic bacteria in the oral cavity. The purpose of this study is to investigate the effect of probiotics isolated from traditional kefir dough on the two main causes of tooth decay.
Methods: Two common species Lactobacillus plantarum and Lactobacillus rhamnosus were confirmed using microbiological techniques (phenotypic and molecular). The antimicrobial effects of these probiotic on Streptococcus mutans (ATCC 35668) and Porphyromonas gingivalis (ATCC 32277) were evaluated using the welling method and then the compounds present in their culture supernatant were identified by gas chromatography Mass spectrometry (GC-MS).
Findings: The experiments revealed that Lactobacillus plantarum and Lactobacillus rhamnosus had a significant antimicrobial effect on Streptococcus mutans and Porphyromonas gingivalis, potentially making them suitable candidates to control the disease. Lactobacillus plantarum decreased gtfB gene expression by 0.001 fold change and brpA gene expression by 0.007 fold change, also Lactobacillus rhamnosus decreased gtfB gene expression by 0.01 fold change and brpA gene expression by 0.04 fold change. Also, GC-MS showed the presence of Pyrrolidione, Furanone, Benzoic acid, Icosane, Nonadecane derivatives in the supernatant of the tested probiotics.
Conclusion: Considering the antimicrobial properties of bacteria isolated from traditional kefir buttermilk, they can be used as traditional probiotic strains with antimicrobial effect in fermented foods.

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Main Subjects


  1. Forssten SD, Björklund M, Ouwehand AC. Streptococcus mutans, caries and simulation models. Nutrients 2010; 2(3): 290-8.
  2. Peres MA, Macpherson LM, Weyant RJ, Daly B, Venturelli R, Mathur MR, et al. Oral diseases: a global public health challenge. Lancet 2019; 394(10194): 249-60.
  3. Ciandrini E, Campana R, Casettari L, Perinelli DR, Fagioli L, Manti A, et al. Characterization of biosurfactants produced by Lactobacillus spp. and their activity against oral streptococci biofilm. Appl
    Microbiol Biotechnol 2016; 100(15): 6767-77.
  4. Evivie SE, Huo GC, Igene JO, Bian X. Some current applications, limitations and future perspectives of lactic acid bacteria as probiotics. Food Nutr Res 2017; 61(1): 1318034.
  5. Khalaf H, Nakka SS, Sandén C, Svärd A, Hultenby K, Scherbak N, et al. Antibacterial effects of Lactobacillus and bacteriocin PLNC8 αβ on the periodontal pathogen Porphyromonas gingivalis. BMC Microbiol 2016; 16(1): 188.
  6. Heymann HO, Swift EJ, Ritter AV. Sturdevant's art
    and science of operative dentistry. 6th Philadelphia, PA: Mosby; 2012.
  7. Alves-Barroco C, Roma-Rodrigues C, Balasubramanian N, Guimarães MA, Ferreira-Carvalho BT, Muthukumaran J, et al. Biofilm development and computational screening for new putative inhibitors of a homolog of the regulatory protein BrpA in Streptococcus dysgalactiae subsp. dysgalactiae. Int J Med Microbiol 2019; 309(3-4): 169-81.
  8. Tahmourespour A, Salehi R, Kermanshahi RK. Lactobacillus acidophilus-derived biosurfactant effect on gtfB and gtfC expression level in Streptococcus mutans biofilm cells. Braz J Microbiol 2011; 42(1): 330-9.
  9. Leite AMO, Miguel MAL, Peixoto RS, Ruas-Madiedo P, Paschoalin VMF, Mayo B, et al. Probiotic potential of selected lactic acid bacteria strains isolated from Brazilian kefir grains. J Dairy Sci 2015; 98(6): 3622-32.
  10. Vos P, Garrity GM, Jones D, Krieg NR, Ludwig W, Rainey FA, et al. Bergey's manual of systematic bacteriology. New York, NY: Springer Science & Business Media; 2011.
  11. Khalaf H, Nakka SS, Sandén C, Svärd A, Hultenby K, Scherbak N, et al. Antibacterial effects of Lactobacillus and bacteriocin PLNC8 αβ on the periodontal pathogen Porphyromonas gingivalis. BMC Microbiol 2016; 16(1): 188.
  12. Jeong D, Kim DH, Song KY, Seo KH. Antimicrobial and anti-biofilm activities of Lactobacillus kefiranofaciens DD2 against oral pathogens. J Oral Microbiol 2018; 10(1): 1472985.
  13. Schleifer KH, Ehrmann M, Beimfohr C, Brockmann E, Ludwig W, Amann R. Application of molecular methods for the classification and identification of lactic acid bacteria. Int Dairy J 1995; 5(8): 1081-94.
  14. Dubernet S, Desmasures N, Guéguen M. A PCR-based method for identification of lactobacilli at the genus level. FEMS Microbiol Lett 2002; 214(2): 271-5.
  15. Gaspar C, Palmeira-de-Oliveira R, Martinez-de-Oliveira J, das Neves J, Pestana PG, Rolo J, et al. Development and validation of a new one step Multiplex-PCR assay for the detection of ten Lactobacillus species. Anaerobe 2019; 59: 192-200.
  16. Hoque MZ, Akter F, Hossain K, Rahman M, Billah M, Islam K. Isolation, identification and analysis of probiotic properties of Lactobacillus spp. from selective regional yoghurts. World J Dairy Food Sci 2010; 5(1): 39-46.
  17. Bhushan J, Chachra S. Probiotics-their role in prevention of dental caries. J Oral Health Comm Dent 2010; 4(3): 78-82.
  18. Ahmed A, Dachang W, Lei Z, Jianjun L, Juanjuan Q, Yi X. Effect of Lactobacillus species on Streptococcus mutans biofilm formation. Pak J Pharm Sci 2014; 27(5 Spec no): 1523-8.
  19. Aboutalebian S, Ahmadikia K, Fakhim H, Chabavizadeh J, Okhovat A, Nikaeen M, et al. Direct detection and identification of the most common bacteria and fungi causing otitis externa by a stepwise multiplex PCR. Front Cell Infect Microbiol 2021; 11: 644060.
  20. Bowen WH, Koo H. Biology of Streptococcus mutans-derived glucosyltransferases: role in extracellular matrix formation of cariogenic biofilms. Caries Res 2011; 45(1): 69-86.
  21. Bitoun JP, Liao S, Yao X, Ahn SJ, Isoda R, Nguyen A, et al. BrpA is involved in regulation of cell envelope stress responses in Streptococcus mutans. Appl Environ Microbiol 2012; 78(8): 2914-22.
  22. Cogulu D, Topaloglu-Ak A, Caglar E, Sandalli N, Karagozlu C, Ersin N, et al. Potential effects of a multistrain probiotic-kefir on salivary Streptococcus mutans and Lactobacillus spp. J Dent Sci 2010; 5(3): 144-9.
  23. Tan YN, Zhang JH, Chen WN. GC-MS-based metabolomics analysis of prawn shell waste co-fermentation by lactobacillus plantarum and bacillus subtilis. Polysaccharides 2020; 1(1): 31-50.
  24. Tahmourespour A, Salehi R, Kermanshahi RK. Lactobacillus acidophilus-derived biosurfactant effect on gtfB and gtfC expression level in Streptococcus mutans biofilm cells. Braz J Microbiol 2011; 42(1): 330-9.