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Original Article

Effectiveness of Vinegar, Lime, and Salt Water as Potential

Household Decontaminants for Toothbrushes
Vethakkan Bijivin Raj, Parangimalai Diwakar Madan Kumar, Selvaraj Balaji1
Departments of Public Health Dentistry, 1Microbiology, Ragas Dental College and Hospital, Chennai, Tamil Nadu, India

Abstract
Introduction: Toothbrushes play an essential role in oral hygiene. However, toothbrushes can become contaminated through contact with the
environment, from the oral cavity, and by toothbrush storage containers. Aim: To evaluate the effectiveness of vinegar, lime, and salt water as
potential household decontaminants for toothbrushes. Materials and Methods: In an in vitro study, 120 used toothbrushes were collected and
divided into four groups comprising 30 samples each. Group I was treated with plain water alone after use. Group II was treated with salt water.
Group III was treated with a solution which contained lime juice. Group IV was treated with vinegar. Treatment duration for the groups was set
to be 12 h. After the treatment, the brush heads of the four groups were incubated in brain-heart infusion agar at 37°C for 24 h. Statistical
analysis was performed using the Statistical Package for the Social Sciences version 20.0 software. One-way analysis of variance and
Bonferroni post hoc analysis were performed for multiple comparisons. Results: Vinegar group showed statistically significant result for
decontamination of toothbrushes when compared to other test and control agents. Conclusion: Commonly used household materials can be
potential decontaminants for toothbrushes. The results of this study showed that vinegar was the most effective decontamination agent
followed by lime and salt water.

Keywords: Antimicrobial, decontamination, household, toothbrush

INTRODUCTION demonstrated that some periodontal pathogenic organisms

and super-infecting Enterobacteriaceae and Pseudomonas
Toothbrushes play a vital role in effective plaque removal and
species were cultured from toothbrushes.[9] Rinsing the
promotion of oral hygiene. Among the seven billion world
toothbrushes using plain water after use may not eliminate
population, 4.2 billion people are using toothbrushes for their
all microorganisms present in the toothbrushes.[10] Hence,
daily oral hygiene maintenance.[1]
studies have been conducted on the potential of various agents
Studies have reported contamination of toothbrushes through for decontamination of toothbrushes.
contact with the environment, from the oral cavity, and by
A number of procedures have been described to reduce the
toothbrush storage containers.[2,3] Dayoub et al.[4] found that
microbiological load of toothbrushes, such as continuous
toothbrushes placed in closed containers and exposed to
replacement of toothbrushes, submerging the brush into
contaminated surfaces of bathrooms yielded higher
microbicide solutions such as Chlorhexidine, sodium
bacterial counts than those left open to air. Mehta et al.[5]
hypochlorite, Listerine, Dettol,[11] spraying antiseptic
found that the use of a cap for toothbrush storage increased
solutions, using ozone, ultraviolet radiation (UV) rays,
bacterial survival. Increased bacterial survival on toothbrush
and microwaves.[10] These procedures are successful in
when stored in moist environment was reported by Glass.[6]
Contaminated toothbrushes can be a vector for the
transmission or reinfection of various bacteria, viruses, Address for correspondence: Dr. V. Bijivin Raj, BDS, Post Graduate Student,
and fungus.[7] Studies stated that Aggregatibacter Department of Public Health Dentistry, Ragas Dental College and Hospital,
actinomycetemcomitans and herpes simplex virus-1 survive 2/102, East Coast Road, Uthandi, Chennai - 600 119, Tamil Nadu, India.
E-mail: bijivin@gmail.com
at least 3 days on toothbrushes.[8] Recent studies
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How to cite this article: Raj VB, Madan Kumar PD, Balaji S.
DOI: Effectiveness of Vinegar, Lime, and Salt Water as Potential
10.4103/jiaphd.jiaphd_120_16 Household Decontaminants for Toothbrushes. J Indian Assoc Public
Health Dent 2017;15:8-10.

8 © 2017 Journal of Indian Association of Public Health Dentistry | Published by Wolters Kluwer - Medknow
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Bijivin Raj, et al.: Household decontaminants for toothbrushes

decontaminating the brushes, but are always expensive or not infusion agar plates were inoculated at 37°C for 24 h. After
easy to perform. Hence, there is a need for a simple and inoculation, total colony count was calculated manually.
inexpensive method for decontaminating toothbrushes. No Total colony count was calculated using the formulae:
study was found to date, which reported decontamination of colony forming unit (cfu)/ml = total colony count × 1000.
toothbrushes using commonly used household materials with
Statistical analysis was performed using IBM Statistical
antibacterial effects.
Package for the Social Sciences Statistics for windows,
Common salt, lime, and vinegar are the most commonly version 20.0 software (IBM Corp., Armonk, NY). Prior to
available household materials with antimicrobial activity. statistical analysis, total microbial counts were converted to
Common salt is an agent used as a food preservative and log10. Descriptive statistics in the form of means and standard
it has high antimicrobial values.[12] Studies have proven lime deviations were calculated. The normality of the data was
juice as a medicine for many infectious diseases, and it can checked using Shapiro–Wilk test, and it was found to be
prevent food contamination.[13] Vinegar is also a household normally distributed. Comparisons of the four groups were
material with antimicrobial activity, and it is also used to conducted with one-way analysis of variance, and Bonferroni
prevent bacterial contamination against food materials.[14] post hoc analysis was performed for multiple comparisons. The
difference was considered statistically significant at P < 0.05.
Considering the availability, cost, and technique sensitivity of
existing household decontamination solutions, this in vitro
study was conceptualized to evaluate the effectiveness of RESULTS
common salt, lime, and vinegar as agents used for The toothbrushes kept in the control group contained more
decontamination of toothbrushes. than 30,000 colonies of microorganisms. Table 1 shows the
distribution of cfu of treatment solutions. Among the four
MATERIALS AND METHODS groups, vinegar showed superior result for decontamination
An in vitro study was conducted in the month of May 2016. The of the toothbrushes when compared to other test and control
toothbrush samples were collected from Panayur, a fishing agents. Vinegar showed a statistically significant reduction in
hamlet near Chennai, India. One hundred and twenty used the microbial count compared to other test solutions among
toothbrushes were collected from people who belong to the age the test agents, which are given in Table 2.
group of 20–30 years with no systemic disease. The study was
explained to them before collecting the toothbrushes and a DISCUSSION
new toothbrush was given to them. They were using these The present study was conducted to analyze the microbial
toothbrushes for around 3 months. After daily use, they rinsed growth after the toothbrush decontamination using household
these brushes using plain water and stored it in their bathrooms agents. The toothbrushes kept in the control group contained
or living rooms. On the day of sample collection after tooth more than 30,000 colonies of microorganisms, which were
brushing in the morning, the samples were collected, and the
head part was separated aseptically using sterile blades and
immersed in the wide-mouthed containers with 5 ml of saline Table 1: Distribution of colony forming unit (cfu) of
and transported to the laboratory on the same day. treatment solutions
The samples were divided into four groups and each group Group Mean ± SD (cfu)
comprised 30 samples. Group I was not treated with any Control 4.2549 ± 0.27751
household agents. They were rinsed with plain water alone Salt water 4.1974 ± 0.16809
after use. This group was considered as control group. Group Lime 3.8281 ± 0.13627
II was treated with salt water. The treatment solution was Vinegar 0.00 (±0.00)
prepared by dissolving two teaspoons of common salt in
100 ml of plain water. Group III was treated with lime juice,
extracted by squeezing the cut, raw lemon. Two teaspoons of Table 2: Bonferroni post hoc test for colony forming unit
lime juice was diluted with 100 ml of plain water. Group IV (cfu) of treatment solutions
was treated with 5 ml of commercially available vinegar (Neo
Groups P value*
Vinegar Company, Chennai, which contains 5–8% of glacial
acetic acid). Treatment duration for the Groups II, III, and IV Control
was set to be 12 h. For all test solutions, care was taken that Salt water 1.000
the toothbrush head was completely submerged in the Lime 0.001
solution. After treatment, the brush heads of the four Vinegar 0.001
groups were transferred to new sterile wide mouth Salt water
containers with 1 ml of saline and subjected to vigorous Lime 0.001
shaking for 5 min. With a 2-mm loop wire, the sample was Vinegar 0.001
Lime
taken from the container and streaked onto brain-heart
Vinegar 0.001
infusion agar (standard loop technique).[15] The brain-heart

Journal of Indian Association of Public Health Dentistry ¦ Volume 15 ¦ Issue 1 ¦ January-March 2017 9
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Bijivin Raj, et al.: Household decontaminants for toothbrushes
in accordance with the existing literature regarding the
toothbrush contamination.[4,9]
Researchers have found that there was an increased risk of
spreading oral and systemic diseases from the contaminated
toothbrushes. Fischer suggested that there was a relationship
between pharyngitis and toothbrush contamination.[16] Brook
and Gober[17] found that the contaminated toothbrushes may
contain group A beta hemolytic streptococci of pharyn-
gotonsillitis. Hence, it is postulated that microbial load in
a toothbrush might have a significant impact in the risk for
systemic diseases.
Many studies have highlighted that there was a need for tooth
disinfection to reduce the number of microorganisms.[3,5,10]
Existing literature shows there are many effective toothbrush
decontamination methods available, which include UV
radiation, microwave radiation,[10] boiling water, and
chemical agents such as Listerine, Plax, Cepacol, and
Chlorhexidine.[18,19] Some authors have also included
agents such as silver and Chlorhexidine to be coated over
the bristles during manufacturing.[19]
Devive et al.[20] quoted that the decontamination method of
the toothbrushes must be rapidly effective, cost effective, and
non-toxic and can be easily implemented.
The present study showed that vinegar had statistically significant
reduction in microbial load. Vinegar contains 5–8% glacial acetic
acid, which is a potent bactericidal agent.[14] It inactivates the
strains of bacteria such as Escherichia coli, thus inhibiting the
growth of microbes. Another test solution, lime juice used in this
study, also provided favorable results. Lime juice contains 8–10%
of citric acid, which is also a bactericidal agent inhibiting the
growth of strains of microorganisms. Salt was a traditionally
used preservative agent which prevents contamination in food
materials.[13] Salt absorbs the moisture content in food materials,
thus preventing bacterial growth and helps to reduce the food
contamination.[12] Our results suggested that vinegar was superior
in decontamination of a toothbrush among household materials
followed by lime and salt water. These results should be
interpreted with the following limitations. In this study,
microbial count of each group was not assessed before the
treatment with various household agents. Furthermore, changes
in the brush bristle pattern could have influenced our research.
No standardized technique of preparation of the solution with
concentration was performed, since it cannot be practiced in a
household setting. In addition, this study aimed to assess
microbial decontamination after 12 h treatment only; hence,
the effect on various time intervals was not assessed. Further
studies may be conducted with lesser treatment time and
microscopic examination of toothbrush bristles after treatment.
CONCLUSION
Commonly used household materials can be potential
decontaminants for toothbrushes. Among the test agents,
vinegar was the most effective decontamination agent
followed by lime and salt water. However, further research
10 Journal of Indian Association of Public Health Dentistry ¦ Volume 15 ¦ Issue 1 ¦ January-March 2017
is recommended on the effect of various concentrations of test
agents at various time intervals.
Financial support and sponsorship
Nil.
Conflicts of interest
There are no conflicts of interest.
REFERENCES
1. Turner J. More-mobile-phones-than-toothbrushes; 2011. [Internet].
Available from: http://60secondmarketer.com/blog. [Last accessed on
2016 Jul 02].
2. ADA.org: ADA statement on toothbrush care: Cleaning, storage and
replacement; 2009. [Internet]. Available from: http://www.ada.org/
1887.aspx. [Last accessed on 2016 Jul 02].
3. Caudry SD, Klitorinos A, Chan EC. Contaminated toothbrushes and
their disinfection. J Can Dent Assoc 1995;61:511-6.
4. Dayoub MB, Rusilko D, Gross A. Microbial contamination of
toothbrushes. J Dent Res 1977;56:706.
5. Mehta A, Sequeira PS, Bhat G. Bacterial contamination and decontami-
nation of toothbrushes after use. N Y State Dent J 2007;73:20-2.
6. Glass RT. The infected toothbrush, the infected denture, and
transmission of disease: A review. Compendium 1992;13:592-8.
7. Glass RT, Carson SR, Barker RL, Peiper SC, Shapiro S. Detection of
HIV proviral DNA on toothbrushes: A preliminary study. J Okla Dent
Assoc 1994;84:17-20.
8. Sconyers JR, Crawford JJ, Moriarty JD. Relationship of bacteremia to
toothbrushing in patients with periodontitis. J Am Dent Assoc
1973;87:616-22.
9. Morris DW, Goldschmidt M, Keene H, Cron SG. Microbial
contamination of power toothbrushes: A comparison of solid-head
versus hollow-head designs. J Dent Hyg 2014;88:237-42.
10. Gujjari SK, Gujjari AK, Patel PV, Subhasini PV. Comparative
evaluation of ultraviolet and microwave sanitization techniques for
toothbrush decontamination. J Int Soc Prev Community Dent
2011;1:20-6.
11. Warren DP, Goldschmidt MC, Thompson MB, Adler-Storthz K, Keene
HJ. The effects of toothpastes on the residual microbial contamination
of toothbrushes. J Am Dent Assoc 2001;132:1241-5.
12. Wijnker JJ, Koop G, Lipman LJ. Antimicrobial properties of salt
(NaCl) used for the preservation of natural casings. Food Microbiol
2006;23:657-62.
13. Hindi NK, Chabuck ZA. Antimicrobial activity of different aqueous
lemon extracts. J Appl Pharm Sci 2013;3:074-8.
14. Medina E, Romero C, Brenes M, De Castro A. Antimicrobial activity
of olive oil, vinegar, and various beverages against foodborne
pathogens. J Food Prot 2007;70:1194-9.
15. Cardoso CL, Muraro CB, Siqueira VL, Guilhermetti M. Simplified
technique for detection of significant bacteriuria by microscopic
examination of urine. J Clin Microbiol 1998;36:820-3.
16. Fischer H. Contaminated toothbrushes and pharyngitis. Arch
Otolaryngol Head Neck Surg 1999;125:479.
17. Brook I, Gober AE. Persistence of group A beta-hemolytic streptococci
in toothbrushes and removable orthodontic appliances following
treatment of pharyngotonsillitis. Arch Otolaryngol Head Neck Surg
1998;124:993-5.
18. Caudry SD, Klitorinos A, Chan EC. Contaminated toothbrushes and
their disinfection. J Can Dent Assoc 1995;61:511-6.
19. Suido H, Offenbacher S, Arnold RR. A clinical study of bacterial
contamination of chlorhexidine-coated filaments of an interdental
brush. J Clin Dent 1998;9:105-9.
20. Devine DA, Percival RS, Wood DJ, Tuthill TJ, Kite P, Killington RA,
et al. Inhibition of biofilms associated with dentures and toothbrushes
by tetrasodium EDTA. J Appl Microbiol 2007;103:2516-24.