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Table of Contents   
ORIGINAL RESEARCH  
Year : 2011  |  Volume : 22  |  Issue : 1  |  Page : 2-5
Assessment of microbial contamination of toothbrush head: An in vitro study


Department of Preventive and Community Dentistry, Bapuji Dental College and Hospital, Davangere, Karnataka, India

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Date of Submission01-May-2010
Date of Decision09-Dec-2010
Date of Acceptance15-Dec-2010
Date of Web Publication25-Apr-2011
 

   Abstract 

Background: The most common oral hygiene aid used to improve the oral health of an individual is toothbrush and it is often neglected to maintain in a proper aseptic condition, which could probably result in contamination by various micro-organisms.
Aims: 1) To assess the microbial contamination of the hard deposit on the toothbrush head, between the bristle tufts, after 1 and 3 months of use. 2) To assess the microbial contamination of the hard deposit on the toothbrush head, between the bristle tufts, which were kept in the bathrooms with and without attached toilet.
Setting and Design: An in vitro study.
Materials and Methods: Twenty toothbrushes kept in the bathrooms with an attached toilet and 20 toothbrushes kept in the bathrooms without an attached toilet were collected from the participants, making it a total of 40. Among these 20 toothbrushes in each group, 10 were used for 1 month duration and 10 were used for 3 months duration.
Results: Streptococcus mutans, Staphylococcus aureus, Pseudomonas, Lactobacillus, Klebsiella, Candida species were isolated in 1 month and 3 months used toothbrushes kept in the bathrooms without attached toilet. Escherichia coli was found in 3 months used toothbrushes kept in the bathrooms with attached toilet. Micro-organisms were found in isolated form in toothbrushes used for 1 month, whereas in toothbrushes used for 3 months they are found in clumps.
Conclusion: Hard deposit on the toothbrush head between bristle tufts is a nidus for growth of micro-organisms, which not only affects the oral health but also affects the general health of an individual.

Keywords: Hard deposit between bristle tufts, microbial contamination, toothbrush contamination

How to cite this article:
Karibasappa G N, Nagesh L, Sujatha B K. Assessment of microbial contamination of toothbrush head: An in vitro study. Indian J Dent Res 2011;22:2-5

How to cite this URL:
Karibasappa G N, Nagesh L, Sujatha B K. Assessment of microbial contamination of toothbrush head: An in vitro study. Indian J Dent Res [serial online] 2011 [cited 2019 Apr 25];22:2-5. Available from: http://www.ijdr.in/text.asp?2011/22/1/2/79965
Oral health is an integral part of general health. It directly and indirectly reflects the overall well-being of an individual, thus maintaining oral hygiene becomes a crucial factor. Oral cavity is free of micro-organisms at birth because the fetus develops in a well-protected environment, but soon after it is habituated by numerous micro-organisms. [1] It may be due to exposure to polluted environment which contains various micro-organisms or change in dietary habits of the child as it grows.

Oral diseases can be greatly controlled by reducing the microbial load in the oral cavity and this can be achieved by maintaining proper oral hygiene. Oral hygiene aids have been in use since Sumerian times. Brushing teeth is the primary mode of oral hygiene practice. In earlier days, chewing sticks like Miswak, Neem and Babul were the sole oral hygiene aids used by different populations. [2] Later, toothbrush crept in as a main component among all oral hygiene aids, as a result of civilization. In 1844, the first toothbrush was manufactured by hand and patented as a three-row brush of serrated bristles with large tufts by Dr. Meyer. L. Rhein. [3]

Tooth brushes are the most commonly used oral hygiene aid [4] to promote oral health and prevent dental diseases. Unfortunately, proper care of toothbrush is often neglected and is kept in bathrooms which are a good place to harbor millions of micro-organisms. The reason attributed to this would be lack of awareness among the public regarding toothbrush maintenance.

Retention and survival of micro-organisms on toothbrush after brushing represents a possible cause of re-contamination of the mouth. [5] Prolonged use of the toothbrush facilitates contamination by various micro-organisms such as Streptococcus, Staphylococcus, [6] and lactobacilli. [7] These micro-organisms are implicated to cause dental caries, gingivitis, [8] stomatitis, infective endocarditis in an individual, affecting both oral and general health. [5]

The average life span of a manual toothbrush is approximately 3 months. [9] Hence, ADA recommends change of toothbrushes once in 3-4 months [10] based on fraying of toothbrush bristles as it decreases the cleaning effectiveness. [10],[11] However, attention was not given to the microbial contamination when the recommendation for frequency of change of toothbrush was given. Many studies have been conducted on toothbrush bristles and anchoring filaments to assess the microbial contamination. However, extensive exploration of the literature reveals lack of studies conducted to assess the microbial contamination of toothbrush head between the bristle tufts. Hence, an in vitro experiment study was designed to assess the microbial contamination of toothbrush head between the bristle tufts.


   Materials and Methods Top


Materials used in the present study included toothbrushes, different agar media, agar plates, plastic sterile container, scissors toothpick, inoculation loops, glucose broth and compound microscope. Permission was obtained from the concerned authorities to carry out the study in the Department of Microbiology, JJM Medical College, and from the Department of Oral pathology and Microbiology, Bapuji Dental College and Hospital, Davangere. Informed consent was obtained from the participants.

Methodology

Forty toothbrushes were collected, 20 toothbrushes in each group based on the key article. [12] Among the 40 toothbrushes, 20 were kept in the bathrooms with an attached toilet and the other 20 were kept in the bathrooms without an attached toilet, after usage. Out of the 20 toothbrushes, 10 were of 1 month duration and the other 10 were of 3 months duration in each group; they were collected from volunteer dental students aged 18-25 years, who were free of any oral and systemic disease. Clinical examination was carried out by the examiner to exclude participants with oral diseases, using mouth mirror, explorer and periodontal probe under aseptic conditions. Medical history of the participants was obtained from the participants themselves to rule out systemic diseases. Later, all the 40 toothbrushes were collected in a plastic sterile container after drying the toothbrushes naturally. Maintaining aseptic measures and adhering to strict protocol, the bristles of toothbrush were trimmed using scissors. With the help of a sterile toothpick, a streak of deposit between the bristle tufts was taken and inoculated into the glucose broth, which supports the growth of micro-organisms and was incubated for 24 hours.

Sterilized agar plates were selected; agar media to allow growth of micro-organisms were prepared by weighing correct proportion of water and agar powder according to manufacturer's instructions and following aseptic protocol. The prepared agar media were poured into the agar plates and kept in the refrigerator to allow cooling of the agar media and to prevent contamination. The hard deposit growth was inoculated into the prepared agar plates with different agar media using inoculation loop. To isolate micro-organisms, various agar media like Mac-Conkey, Blood, Mitis Salivarius Bacitracin, Rogosa, and Sabourauds were used. After inoculation, agar plates were kept in an incubator to allow for the growth of the micro-organisms. Standardization was done with respect to materials, instruments, methodology and calibration for the microbial analysis. After 24 hours, agar plates were observed and examined using compound microscope. As the data obtained were nominal in nature, the present study was not subjected to statistical analysis.


   Results Top


Microbiological analysis using compound microscope revealed the following micro-organisms in 1 month and 3 months used toothbrushes kept in the bathrooms with and without attached toilets.

[Table 1] shows micro-organisms found in 1 month used toothbrushes kept in the bathroom with and without attached toilet. In the majority of tooth brushes kept in bathroom without attached toilet, micro-organisms like Streptococcus mutans, Candida, Pseudomonas, and Klebsiella were seen. Staphylococcus aureus Scientific Name Search  was seen in addition to these micro-organisms in 1 month used toothbrushes kept in the bathroom with attached toilet.
Table 1: Micro-organisms in 1 month used toothbrushes kept in the bathroom with and without attached toilet

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[Table 2] shows micro-organisms found in 3 months used toothbrushes kept in the bathroom with and without attached toilet. Majority of the tooth brushes were contaminated with microorganisms like Str. mutans, Pseudomonas, and Streptococcus sanguis.  Escherichia More Details coli was seen in addition to these micro-organisms in 3 months used toothbrushes kept in the bathroom with attached toilet.
Table 2: Micro-organisms in 3 months used toothbrushes kept in the bathroom with and without attached toilet

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   Discussion Top


In the present study, toothbrush head between the bristle tufts was selected to assess the microbial contamination as it not only harbors micro-organisms but also provides a favorable environment for their growth. The contamination of tooth brush could pose a significant risk of dissemination of infection for certain patients such as immunosuppressed, cardiopathic, organ transplant recipients. [13] Micro-organisms were assessed in 1 month and 3 months used toothbrushes as 1 month used toothbrush had sufficient hard deposit to facilitate isolation of it, and according to ADA recommendations toothbrushes have to be changed once in 3-4 months. [10]

This study revealed micro-organisms to be present in the toothbrush head between the bristle tufts. Micro-organisms were found in isolated form and were less in number in 1 month used toothbrush, whereas they were in clumps and more in number in 3 months used toothbrushes. The micro-organisms which were found in 1 month used toothbrush kept in the bathroom included Str. mutans, Lactobacilli, Candida, Pseudomonas, Klebsiella, Str. pyogenes, Sta. aureus [Table 1]. This result is in agreement with the study conducted by Willi-Eckhard Wetzel et al. in the year 2005. [5]

Various micro-organisms isolated in this study cause different diseases, e.g., Str. mutans causes initiation of dental caries in human beings; Lactobacilli cause the progression of the dental caries; Candida causes candidiasis; Pseudomonas causes suppurative otitis, eye infections, urinary tract infections, burn infections, etc.; Klebsiella causes pyogenic infections, septicemia, pneumonia, diarrhea, urinary tract infections; Str. pyogenes causes urinary tract infections, rheumatic fever, glomerulonephritis; and Sta. aureus causes boils, carbuncle, pustules, abscess, osteomyelitis, endocarditis and septicemia.

The micro-organisms in 3 months used toothbrush kept in bathrooms with attached toilet included E. coli apart from the organisms which were found in 1 month used toothbrushes kept in the bathroom with or without attached toilet and 3 months used toothbrushes kept in the bathroom without attached toilet [Table 2]. This is in agreement with the study conducted by Suma Sogi et al.[13] E. coli is known to cause diarrhea, urinary tract infections, septicemia. Micro-organisms were found to be present in all the toothbrushes except two which were used for 1 month and kept in the bathrooms with and without attached toilet. This may be due to failure in the hard deposit collection or failure in inoculation into agar plates.

In the present study, the micro-organisms isolated were not only oral pathogens but also general pathogens. The source of general pathogens could be improper storage of toothbrushes that were kept in the bathrooms with or without attached toilet, exposing them to the unfavorable surrounding external environment. [14] Oral commensals could also have contributed for contamination of toothbrushes. Toothbrushes contaminated with the micro-organisms which are potent pathogens like Pseudomonas, Sta. aureus and Klebseilla will definitely pose a serious threat to oral as well as general health.

Limitations of the current study

As microbial growth occurred in clumps, colony counting was not possible and therefore the level of pathogenicity was not known. Toothbrushing with contaminated toothbrushes might lead to development of certain diseases depending upon the type of micro-organisms present on the toothbrushes. In the present study, only healthy volunteers free of oral and systemic diseases were selected. However, occurrence of disease in the participants due to toothbrushing with contaminated toothbrushes remained unknown. In addition to this, only toothbrushes used for 1 month and 3 months duration were examined to know their microbial contamination.

Further studies have to be conducted to assess the microbial contamination of toothbrush head used for 15 days, 1.5, 2, and 2.5 months, to know the pathogenicity of micro-organisms and to evaluate the occurrence of any infection in the subjects following the use of contaminated toothbrushes.

Several methods are available to preserve toothbrushes like usage of anti-microbial solutions, natural air drying and toothbrush sanitizer. Some of the commercially available antimicrobial solutions are 0.2% Chlorhexidine, 2% Triclosan, 1% Sodium hypochlorite, [15] 3% Hydrogen peroxide, Dettolin, [13] etc. Home-made microbial solutions like 3% neem [15] that has antimicrobial property equivalent to that of standard antimicrobial solutions can also be recommended. The efficacy of salt water as an anti-microbial agent for preservation of toothbrushes should be tested and, if proved efficient, could be recommended.


   Conclusions Top


Frequency of change of toothbrushes based on fraying alone should not be considered. Whenever hard deposits are appreciated on the head of the toothbrush, it should be viewed as a nidus for microbial contamination. Ideally, toothbrushes should not be kept in the bathroom with and without attached toilets as it is prone for contamination. Otherwise, toothbrushes should be kept in the anti-microbial solutions to prevent microbial contamination.


   Acknowledgments Top


The authors acknowledge the help of Dr. Vishwanath, Professor and head, Department of Microbiology, JJM Medical College and hospital, Davangere, and Dr. Mujib Ahmed, Professor and Head, Department of Oral Pathology and Oral Microbiology, Bapuji Dental College and Hospital, Davangere.

 
   References Top

1.McCarthy C, Synder ML, Parker RP. The indigenous oral flora of man. The newborn to the 1 year old infant. Arch Oral Biol 1965;10:61-70.  Back to cited text no. 1
    
2.Bhat S, Hegde KS, George R. Microbial contamination of toothbrushes and their decontamination. J Indian Soc Pedo Prev Dent 2003;21:108-12.  Back to cited text no. 2
    
3.Toothbrush origin - Toothbrush history. Available from: http://www.toothbrush express.com/html/toothbrush_history.html [last accessed on 2010 Apr 20].  Back to cited text no. 3
    
4.Carranza FA Jr, Newman MG. A textbook of Clinical Periodontology. 8th ed. Philadelphia: W. B. Saunders; 1996. p. 493.  Back to cited text no. 4
    
5.Wetzel WE, Schaumburg C, Ansari F, Kroeger T, Sziegoleit A. Microbial contamination of toothbrushes with different principles of filament anchoring J Am Dent Assoc 2005;136:758-64.  Back to cited text no. 5
    
6.Taji SS, Rogers AH. The microbial contamination of toothbrush: A pilot study. Aust Dent J 1998;43:128-30.  Back to cited text no. 6
    
7.Fernandez V, Cesar D. Microbiology evaluation of toothbrushes. In Vitro Cell Dev Biol Anim 2006;42:31A.  Back to cited text no. 7
    
8.Boylan R, Li Y, Simeonova L, Sherwin G, Kreismann J, Craig RG, et al. Reduction in bacterial contamination of toothbrushes using violight, ultraviolet light activated toothbrush sanitizer. Am J Dent 2008;21:313-7.  Back to cited text no. 8
    
9.Yankell SL, Saxer UP. Toothbrushes and Toothbrushing methods. In: Harris NO, Garcia-Godoy F. Primary Preventive Dentistry. 6th ed. New Jersy: Prentice Hall; 2004. p.110.  Back to cited text no. 9
    
10.ADA: Time to change that brush. Chain Drug Review 2009. Available from: http://www.findarticles.com/p/articles/mi_hb3007/is_3_31/ai_n31352565/ [last accessed on 2010 Apr 20].  Back to cited text no. 10
    
11.Glaze PM, Wade AB. Toothbrush age and wear as it relates to plaque control. J Clin Periodontol 1986;13:52-6.  Back to cited text no. 11
    
12.Saravia ME, Nelson-Filho P, da Silva RA, Faria G, Rossi MA, Ito IY. Viability of Streptococcus mutans toothbrush bristles. ASDC J Dent Child 2008;75:29-32.  Back to cited text no. 12
    
13.Sumasogi HP, Subbareddy VV, ShashiKiran ND. Contamination of toothbrush at different time intervals and effectiveness of various disinfecting solutions in reducing the contamination of toothbrush. J Indian Soc Prev Dent 2002;20:81-5.   Back to cited text no. 13
    
14.Long SR, Santos AS, Nascimento CM. Avaliação da contaminação de escovas dentais por enterobactérias. Rev Odontol Univ Santo Amaro 2000;5:21-5.   Back to cited text no. 14
    
15.Balappanavar AY, Nagesh L, Ankola AV. Antimicrobial efficacy of various disinfection solutions in reducing the contamination of toothbrushes. Oral Health Prev Dent 2009;7:137-45.  Back to cited text no. 15
    

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Correspondence Address:
G N Karibasappa
Department of Preventive and Community Dentistry, Bapuji Dental College and Hospital, Davangere, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0970-9290.79965

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    Tables

  [Table 1], [Table 2]

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