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Year : 2013  |  Volume : 1  |  Issue : 3  |  Page : 95-98

Antimicrobial effect of different xylitol concentrations on Streptococcus mutans and Lactobacillus acidophilus count


Department of Restorative Dentistry, College of Dentistry, University of the East, Sampaloc, Manila, Philippines

Date of Web Publication25-Sep-2013

Correspondence Address:
S Radmerikhi
Department of Restorative Dentistry, College of Dentistry, University of the East, Sampaloc, Manila
Philippines
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2321-4619.118907

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  Abstract 

In recent years, pentacarbon sugars including xylitol are employed as supplements in the preparation of oral health products. The main purpose of this study was to measure Streptococcus mutans and Lactobacillus acidophilus count treated with different xylitol concentrations. Bacterial solutions were mixed separately in to 2 M (30.43%) xylitol stock solution. The mixture provided bacterial growth medium with 3-18% of xylitol. The solutions stored in a 37°C incubator for 48 h. Initial optical density and optical density of samples after 48 h taken using spectrophotometer. Result shown in average, bacterial growth in L. acidophilus was higher than S. mutans and even in higher xylitol concentration this difference preserved. The present study indicates that the addition of xylitol drastically can enhance the effect of antibacterial agent.

Keywords: Antimicrobial effect, Lactobacillus acidophilu, Streptococcus mutans, xylitol


How to cite this article:
Radmerikhi S, Formantes B, Fajardo K R, Azul E. Antimicrobial effect of different xylitol concentrations on Streptococcus mutans and Lactobacillus acidophilus count. J Res Dent 2013;1:95-8

How to cite this URL:
Radmerikhi S, Formantes B, Fajardo K R, Azul E. Antimicrobial effect of different xylitol concentrations on Streptococcus mutans and Lactobacillus acidophilus count. J Res Dent [serial online] 2013 [cited 2019 Dec 14];1:95-8. Available from: http://www.jresdent.org/text.asp?2013/1/3/95/118907


  Introduction Top


Two groups of bacteria are responsible for initiating caries: Streptococcus mutans[1] and Lactobacillus acidophilus.[2] If left untreated, the disease can lead to pain, tooth loss and infection. Tooth decay is caused by specific types of bacteria that produce acid in the presence of fermentable carbohydrates such as sucrose, fructose, and glucose. [3],[4] In fact the mineral content of teeth is sensitive to increases in acidity from the production of lactic acid. In recent years, pentacarbon sugars, pentiols, including xylitol, are employed as supplements in the preparation of oral hygiene products. Xylitol may not only improve taste value of the toothpaste but may also the environment of the oral cavity. [5] Xylitol is a 5-carbon polyol sugar alcohol commonly used as a non-cariogenic sweetener in chewing gums, tablets, dentifrice, and oral rinses. [6] Previous studies have demonstrated the cariostatic properties of xylitol, which may be linked to the inhibited development of dental plaque and to the antibacterial activity toward S. mutans and L. acidophilus, microbes responsible for development of the decay process. [5] Scheinin et al. [7] have noted a significantly reduced occurance of teeth decay by use of a xylitol containing toothpaste, but Twetman and Peterson failed to confirm the effect. The use of high concentrations of xylitol have been shown to inhibit Lactococcus lactis Scientific Name Search  over time, but not on Streptococcus salivarius and L. casei.[8] Further, xylitol has been shown to prevent the adherence of Pneumococci and Haemophilus influenzae to nasopharyngeal cells due to a fructose phosphotransferase system-mediated uptake and phosphorylation of xylitol in the cell. [4],[9],[10] Studies have demonstrated growth inhibition for Streptococcus mitis with xylitol but not in Streptococcus milleri was not inhibited by it. [11],[12] In vitro studies support this idea; Actinomyces strains, Vitellariopsis dispar and Fusobacterium nucleatum were not inhibited by xylitol. [13] S. mutans has been implicated as a principal etiological agent of dental caries in humans because this oral bacterium has the ability to adhere to and produce acid at the tooth surface when carbohydrates, sucrose, serve as a substrate. [14] The aim of this study was to determine and compare, S. mutans and L. acidophilus population change when treated with 3%, 8%, 12% and 18% xylitol concentrations.


  Materials and Methods Top


Bacterial strains

Freeze dried S. mutans and L. acidophilus were obtained from Philippine National Collection of Microorganisms (PNCM), University of the Philippines, Los Baρos, College, Laguna, Philippines, together with transfer and growth media. The cells were re-suspend with 0.2 ml of the liquid transfer medium provided for each strains.

Preparation of xylitol stock solution

30.43% or 2M xylitol stock solution was prepared by dissolving 30.43 g xylitol in 100 ml distilled water. 30.43% aqueous xylitol solution was made by dissolving 30 g of xylitol in 100 ml distilled water. The stock solution was autoclaved at 121°C for 15 min.

Bacterial transfer

2.72 ml, 2.22 ml, 1.83 ml and 1.23 ml of S. mutans and L. acidophilus acidophilus cultures were added in 0.29 ml, 0.78 ml, 1.17 ml and 1.77 ml of 2M (30.43%) xylitol stock solution. The mixture provided 3 ml of bacterial growth medium with 3%, 8%, 12% and 18% of xylitol. Likewise, 3 ml of bacteria in broth was prepared to serve as control. All the solutions were stored in a 37°C incubator.

Antimicrobial assay using spectrophotometer

Initial optical density and optical density of samples after 48 h were taken using spectrophotometer. Optical density of samples read at 600 nm to measure optical density of S. mutans[15] and L. acidophilus[16] by using spectrophotometer (Stat Fax® 4500). All the experiments were performed in three trials.

Statistical analysis

Spectrophotometry result were subjected to two-way analysis of variance (ANOVA) test followed by Tukey's multiples comparison test using IBM SPSS Statistics v. 20 software. Diagrams were prepared using Microsoft Excel 2013.


  Results Top


Analysis of results using ANOVA showed a significant difference in the optical density between S. mutans and L. acidophilus in different xylitol concentrations [Table 1]. Result shown by an increase in xylitol concentration mean for both microorganisms, lower optical density was observed with higher xylitol concentrations. Marked inhibition of bacterial growth occurred in 18% xylitol concentration [Table 2], [Figure 1] and [Figure 2].
Figure 1 : Effect of different xylitol concentration on Streptococcus mutans

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Figure 2: Effect of different xylitol concentration on Lactobacillus acidophilus

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Table 1: Result of ANOVA analysis

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Table 2: Mean of bacterial growth in different concentration

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Result shown in average, bacterial growth in L. acidophilus (2.038) was higher than S. mutans (1.059 OD) and even in higher xylitol concentration this difference preserved [Table 3] and [Table 4], [Figure 3].
Figure 3: Effect of different xylitol concentration on Streptococcus mutans

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Table 3: Mean of bacterial growth

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Table 4: Mean comparison of bacterial growth in different concentration

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Result shown highest optical density in medium with S. mutans belong to control group (1.484 OD), lowest optical density in medium with S. mutans belong to medium with 18% xylitol (0.688) and optical density decrease 0.797 OD by an increase in xylitol concentration up to 18%. Also, the result shows that the highest optical density in medium with L. acidophilus belongs to control group (2.496 OD), lowest optical density in medium with L. acidophlus belongs to medium with 18% xylitol (1.434) and optical density decrease 1.062 OD by an increase in xylitol concentration up to 18%. Result shown in both bacteria by an increase in xylitol concentration the bacterial count decreased by half [Table 4].

Result of Tukey's multiple test shown 99% confidence of significant difference between 8%, 12% and 18% with other groups and also result shown 95% confidence of significant difference between 3% xylitol and control group.


  Discussion Top


The present study indicates that the addition of xylitol drastically can enhance the effect of antibacterial agent. Also, the study demonstrated that xylitol consumption affect the composition of oral flora by a decrease in S. mutans and L. acidophilus. Xylitol inhibits the growth of bacteria via the inducible fructose phosphptrasnferase system and formation xylitol-5-phosphate. Also, subsequent accumulation of xylitol-5-phosphate interferese with carbohydrate methabolism. [17],[18] Also it can disturbs the protein synthesis and causes ultrastructural changes bacteria. [10],[19] Subsequent accumulation of xylitol Result of this study confirms that S. mutans and L. acidophilus, also possess the fructose pathway and could thus be inhibited by xylitol as pervious report [20] on L. paracasei, L. brevis and L. fermentum. The effect of xylitol on bacterial level depend to both of, its high frequency and the concentration of the agent. [21],[22] Although previous studies shown other organism are also sensitive to xylitol, but the decrease in bacterial level should be more intense on S. mutans and L. acidophilus due to decrease in the period of acid pH in the dental plaque. [23],[24],[25]


  Conclusion Top


Our result suggest that the consumption of 5% xylitol reduced S. mutans and L. acidophilus growth and by an increase in xylitol inhibitory effect increased. This study confirms that even in low concentrations, Xylitol has an inhibitory effect against main oral cavity inducer bacteria. Xylitol has more inhibitory effect on L. acidophilus in comparison with S. mutans. Further studies on long time effect of the xylitol solution from 8 to 12 week and effect of very low concentration of xylitol against S. mutans and L. acidophilus is recommending.


  Acknowledgment Top


This research is a portion of the thesis submitted in partial fulfillment of the requirements of the degree of Doctor of Dental Medicine in the University of the East - College of Dentistry. The authors would like to thank Dr. H. Ahmadian Moghadam for her patience, encouragement and insightful comments.

 
  References Top

1.Holbrook WP, Magnusdottir MO. Studies on strains of Streptococcus mutans isolated from caries-active and caries-free individuals in Iceland. J Oral Microbiol 2012;4.  Back to cited text no. 1
    
2.Jay P. Lactobacillus Acidophilus and Dental Caries. Am J Public Health Nations Health 1938;28:759-61.  Back to cited text no. 2
    
3.Hardie JM. The microbiology of dental caries. Dent Update 1982;9:199-200, 202-4, 206-8.  Back to cited text no. 3
    
4.Kontiokari T, Uhari M, Koskela M. Antiadhesive effects of xylitol on otopathogenic bacteria. J Antimicrob Chemother 1998;41:563-5.  Back to cited text no. 4
    
5.Assev S, Waler SM, Rolla G. Xylitol fermentation by human dental plaque. Eur J Oral Sci 1996;104:359-62.  Back to cited text no. 5
    
6.Twetman S, Petersson LG. Influence of xylitol in dentifrice on salivary microflora of preschool children at caries risk. Swed Dent J 1995;19:103-8.  Back to cited text no. 6
    
7.Scheinin A, Banoczy J, Szoke J, Esztari I, Pienihakkinen K, Scheinin U, et al. Collaborative WHO xylitol field studies in Hungary. I. Three-year caries activity in institutionalized children. Acta Odontol Scand 1985;43:327-47.  Back to cited text no. 7
    
8.Walters La. The Effect of xylitol on the growth of three normal oral commensal or probiotic Bacteria [Bachelor of Science]. Coastal Carolina University, 2011.  Back to cited text no. 8
    
9.Tapiainen T, Kontiokari T, Sammalkivi L, Ikaheimo I, Koskela M, Uhari M. Effect of xylitol on growth of Streptococcus pneumoniae in the presence of fructose and sorbitol. Antimicrob Agents Chemother 2001;45:166-9.  Back to cited text no. 9
    
10.Trahan L, Bareil M, Gauthier L, Vadeboncoeur C. Transport and phosphorylation of xylitol by a fructose phosphotransferase system in Streptococcus mutans. Caries Res 1985;19:53-63.  Back to cited text no. 10
    
11.Kontiokari T, Uhari M, Koskela M. Effect of xylitol on growth of nasopharyngeal bacteria in vitro. Antimicrob Agents Chemother 1995;39:1820-3.  Back to cited text no. 11
    
12.Rogers AH, Pilowsky KA, Zilm PS, Gully NJ. Effects of pulsing with xylitol on mixed continuous cultures of oral streptococci. Aust Dent J 1991;36:231-5.  Back to cited text no. 12
    
13.Bradshaw DJ, Marsh PD. Effect of sugar alcohols on the composition and metabolism of a mixed culture of oral bacteria grown in a chemostat. Caries Res 1994;28:251-6.  Back to cited text no. 13
    
14.Gibbons RJ, van Houte J. Dental caries. Annu Rev Med 1975;26:121-36.  Back to cited text no. 14
    
15.Michalek SM, McGhee JR, Babb JL. Effective immunity to dental caries: Dose-dependent studies of secretory immunity by oral administration of Streptococcus mutans to rats. Infect Immun 1978;19:217-24.  Back to cited text no. 15
    
16.Goderska K, Nowak J, Czarnecki Z. Comparison of the growth of Lactobacillus acidophilus and Bifidobacterium bifidum species in media supplemented with selected saccharides including prebiotics. Acta Sci Pol Technol Aliment 2008;7:5-20.  Back to cited text no. 16
    
17.Miyasawa-Hori H, Aizawa S, Takahashi N. Difference in the xylitol sensitivity of acid production among Streptococcus mutans strains and the biochemical mechanism. Oral Microbiol Immunol 2006;21:201-5.  Back to cited text no. 17
    
18.Giertsen E, Emberland H, Scheie AA. Effects of mouth rinses with xylitol and fluoride on dental plaque and saliva. Caries Res 1999;33:23-31.  Back to cited text no. 18
    
19.Tuompo H, Meurman JH, Lounatmaa K, Linkola J. Effect of xylitol and other carbon sources on the cell wall of Streptococcus mutans. Scand J Dent Res 1983;91:17-25.  Back to cited text no. 19
    
20.Helanto M, Aarnikunnas J, Palva A, Leisola M, Nyyssola A. Characterization of genes involved in fructose utilization by Lactobacillus fermentum. Arch Microbiol 2006;186:51-9.  Back to cited text no. 20
    
21.Lif Holgerson P, Stecksen-Blicks C, Sjostrom I, Twetman S. Effect of xylitol-containing chewing gums on interdental plaque-pH in habitual xylitol consumers. Acta Odontol Scand 2005;63:233-8.  Back to cited text no. 21
    
22.Ly KA, Milgrom P, Roberts MC, Yamaguchi DK, Rothen M, Mueller G. Linear response of mutans streptococci to increasing frequency of xylitol chewing gum use: A randomized controlled trial [ISRCTN43479664]. BMC Oral Health 2006;6:6.  Back to cited text no. 22
    
23.Caglar E, Kavaloglu SC, Kuscu OO, Sandalli N, Holgerson PL, Twetman S. Effect of chewing gums containing xylitol or probiotic bacteria on salivary mutans streptococci and lactobacilli. Clin Oral Investig 2007;11:425-9.  Back to cited text no. 23
    
24.Decker EM, Maier G, Axmann D, Brecx M, von Ohle C. Effect of xylitol/chlorhexidine versus xylitol or chlorhexidine as single rinses on initial biofilm formation of cariogenic streptococci. Quintessence Int 2008;39:17-22.  Back to cited text no. 24
    
25.Holgerson PL, Sjostrom I, Twetman S. Decreased salivary uptake of [14C]-xylitol after a four-week xylitol chewing gum regimen. Oral Health Prev Dent 2007;5:313-9.  Back to cited text no. 25
    


    Figures

  [Figure 1], [Figure 2], [Figure 3]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4]


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