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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 is recommended on the effect of various concentrations of test
toothbrush contamination.[4,9] agents at various time intervals.
Researchers have found that there was an increased risk of
Financial support and sponsorship
spreading oral and systemic diseases from the contaminated
toothbrushes. Fischer suggested that there was a relationship Nil.
between pharyngitis and toothbrush contamination.[16] Brook
and Gober[17] found that the contaminated toothbrushes may Conflicts of interest
contain group A beta hemolytic streptococci of pharyn- There are no conflicts of interest.
gotonsillitis. Hence, it is postulated that microbial load in
a toothbrush might have a significant impact in the risk for REFERENCES
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10 Journal of Indian Association of Public Health Dentistry ¦ Volume 15 ¦ Issue 1 ¦ January-March 2017