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ORIGINAL ARTICLE
Year : 2014  |  Volume : 34  |  Issue : 2  |  Page : 85-88

Efficacy of garlic extract and chlorhexidine mouthwash in reduction of oral salivary microorganisms, an in vitro study


Department of Paedodontics and Preventive Dentistry, Yenepoya Dental College, Yenepoya University, Deralakatte, Mangalore, Karnataka, India

Date of Web Publication18-Mar-2015

Correspondence Address:
H T Ajay Rao
Department of Paedodontics and Preventive Dentistry, Yenepoya Dental College, Yenepoya University, University Road, Nithyananda Nagar Post, Deralakatte, Mangalore - 575 018, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0257-7941.153465

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  Abstract 

Objectives: To assess and compare the antimicrobial effect of garlic extract and chlorhexidine (CHX) mouthwash solution against oral salivary microorganisms.
Materials and Methods: Thirty six salivary samples were obtained in dry plastic vials. Collected saliva samples were centrifuged. Each vial was mixed completely on a shaker after which 1 ml of saliva was added to 9 ml of ethanol by a sterile pipette and mixed. A volume of 1 ml of garlic hydro-alcoholic extract and 1 ml of mouthwash was added to 1 ml each case specimen and was transferred to culture medium of Trypticase Soy Agar. Agar plates were incubated at 37°C for 48 h to allow for microbial growth. Microbial colonies were counted by independent interpreter to evaluate the result.
Statistical Analysis: Data obtained were analyzed using one-way ANOVA test. P < 0.001 was considered statistically significant.
Result: Result of the study shows that mean colony count of salivary microbial population was (1984 ± 400) 1127 in saline group (negative control), (50 ± 4) 27 in (0.12%) CHX group (positive control), (700 ± 200) 469 in garlic extract (5%) group (case control).
Conclusion: Mouthwash containing garlic extract can be used as an alternative to CHX mouthwash.

Keywords: Chlorhexidine mouthwash, garlic extract, oral microorganisms


How to cite this article:
Ajay Rao H T, Bhat SS, Hegde S, Jhamb V. Efficacy of garlic extract and chlorhexidine mouthwash in reduction of oral salivary microorganisms, an in vitro study. Ancient Sci Life 2014;34:85-8

How to cite this URL:
Ajay Rao H T, Bhat SS, Hegde S, Jhamb V. Efficacy of garlic extract and chlorhexidine mouthwash in reduction of oral salivary microorganisms, an in vitro study. Ancient Sci Life [serial online] 2014 [cited 2020 Sep 26];34:85-8. Available from: http://www.ancientscienceoflife.org/text.asp?2014/34/2/85/153465


  Introduction Top


The living human body consists of innumerable channels called srotas. Ayurveda advocates samśodhana to be the way to clean these channels in order to enable self-healing. Samśodhana as a process also enables the medicines reach the target sites more easily. Samśodhana is considered a prerequisite before administration of any medications and therapeutic interventions. [1] Plants and natural products can treat bacterial infections. Plants are rich in a wide variety of secondary metabolites, such as tannins, terpenoids, alkaloids, and flavonoids, which have been found in vitro to have antimicrobial properties. [2] Most of the oral diseases are due to bacterial infections, and it has been well documented that medicinal plants exhibit considerable antibacterial activity against various microorganisms including the bacteria responsible for dental caries. [3] Hundreds of plants are used in traditional medicine as a treatment for bacterial infections. Garlic (Allium sativum) is one of the medicinal plants that possess sulphides that has antimicrobial properties. [4] Garlic extract has an inhibitory effect on the growth of various bacteria such as Pseudomonas aeruginosa, and Mycobacterium tuberculosis. [5] On the other hand, commercially available antibacterial agents such as chlorhexidine (CHX) have been used in the prevention and management of oral diseases but these agents increase the level of dental staining. [6] Thus, the purpose of the present study was to evaluate the antimicrobial efficacy of 5% garlic extract and compare with 0.12% CHX. Garlic has good antimicrobial activity against periodontal pathogens like Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans and has therapeutic value in periodontitis and in oral infections. [7]


  Materials and Methods Top


Inclusion criteria

Patients requiring oral prophylaxis and restorative treatment.

Exclusion criteria

Patients who had consumed antibiotics in past month before the onset of the study. Patient with systemic diseases.

Saliva sample collection

Twelve patients were selected to participate in this "in vitro" experimental study. Saliva samples were collected from all 12 subjects in dry plastic vials by spitting. Each salivary sample was divided into three groups, case control, positive control and negative control (total samples 36).

Garlic extract preparation

Fresh garlic cloves were washed, peeled and cut into pieces and grinded in grinder then it was added to 95% ethanol. After adding into ethanol, it kept into rotary shaker for 24 h so that all contents of garlic mix into ethanol after that it kept into a rotary evaporator for evaporation of all the ethanol. After evaporation garlic extract was collected then 5 mg this garlic extract was added into 100 ml of saline to make 5% w/v concentration of garlic extract. This method of preparation of the extract is known as hydro-alcoholic extraction method that was also used in the study done by Borhan-Mojabi et al. [8] This 5% of garlic extract was used in the present study [Figure 1]a and b.
Figure 1: (a) Raw garlic extract. (b) Filter sterilized aqueous extract of garlic. (c) Agar plates

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Method

A volume of 1 ml of each of the salivary sample was added to 1 ml of (5%) garlic hydro-alcoholic extract (case control), 1 ml of (0.12%) CHX (positive control), and 1 ml of normal saline (negative control). They were transferred to the culture medium of Trypticase Soy Agar for 30 s. Agar plates were incubated at 37°C for 24 h to allow for microbial growth [Figure 1]c. Microbial colonies were counted by independent interpreter to evaluate the result.

Statistical analysis

Data were evaluated between the groups by one-way ANOVA test. P < 0.001 was considered as statistically significant [Table 1] and [Table 2].
Table 1: Comparison of the colony count using one-way ANOVA


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Table 2: Multiple comparison between groups


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  Result Top


Mean colony count of salivary microbial population was (1984 ± 400) 1127 in saline group (negative control) 1984 is highest number of microorganisms found in saline group out of 12 samples and 400 is lowest number of microorganisms found in saline group out of 12 samples and mean value of microorganisms were 1127 in the saline group [Figure 2]a, (50 ± 4) 27 in (0.12%) CHX mouthwash group (positive control) 50 is the highest number of microorganisms found in CHX group out of 12 samples and 4 is lowest number of microorganisms found in CHX group out of 12 samples and mean value of microorganisms were 27 in the CHX group [Figure 2]b, (700 ± 200) 469 in garlic extract (5%) group (case control) 700 is highest number of microorganisms found in garlic extract group out of 12 samples and 400 is lowest number of microorganisms found in garlic extract group out of 12 samples and mean value of microorganisms were 469 in the garlic extract group [Figure 2]c and Graph 1].
Figure 2: (a) Salivary microbial population in saline group. (b) Salivary microbial population in chlorhexidine mouthwash group. (c) Salivary microbial population in garlic extract group

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  Discussion Top


Herbal extracts have been successfully used in dentistry as tooth cleaning and for plaque reduction. Natural phytochemicals can be an effective alternative to antibiotics and represent a promising approach in the prevention of dental caries and other oral infection. [3] The relationship between oral diseases and oral microbiota is well recognized. Natural plants that have been used as traditional medicines are regarded as good choices. [8] Ayurvedic name of garlic is rasona. According to texts Caraka samhitā, Suśruta samhitā, Rāja nighaṇ ṭu and Bhāvaprakāśa, rasona is a nourishing tonic for the whole body, age sustainer, immune booster, strength promoter, and to be good for complexion and eyesight. [9] Garlic (A. sativum) has been used as a medicine since ancient times and has long been known to have antibacterial, antifungal and antiviral properties. [10] Garlic has been described to have madhura (sweet), lavaṇa, kaṭu, kaṣāya , rasas (tastes). Its guṇas (special properties of the medicinal herb) have been described to be: Snigdha (unctuous), tīkṣṇa (sharp), picchila (slimy), guru (heavy), sara (mobile). The vipāka of garlic is kaṭu (pungent) , and its vīrya (potency) has been described to be uṣ ṇ a (hot). [11] Garlic is also effective against antibiotic resistant organisms [12] and has the ability to kill all oral streptococci strains. [13] Garlic extract has antimicrobial activity against oral bacterial species, particularly Gram-negative species. Its action against P. gingivalis and the allicin present in garlic indicates therapeutic use for periodontitis and possibly other oral infections. Allicin was also found effective against both germination of spores and growth of hyphae [14] it is considered to be most therapeutically potent constituent of garlic. Traditional medicinal use of garlic suggests the use of allicin for alleviating or decreasing the dental diseases. [15] Considering that garlic is essentially a collection of numerous properties that are all beneficial in the management of oral microbial diseases, it may be considered an alternative or supplementary medicine for patients with this disease. Elnima et al. compared the aqueous extracts of garlic (A. sativum) and onion (Allium cepa) for activity against Gram-positive organisms, Gram-negative organisms and fungi and showed that garlic extract had greater activity as compared to the extract of onion. A mouthwash containing 10% garlic in quarter Ringer solution produced a drastic reduction in the number of oral bacteria. [16] In the present study, there was a definitve reduction in oral salivary microbial population in in the sample with 5% garlic extract when exposed for 30 s. The result of our study was comparable with the study done by Borhan-Mojabi et al. [8] In addition, since there is no evidence for severe adverse effects by the use of higher concentrations of garlic extracts, they may be less harmful for patients compared to other antimicrobial drugs with similar effects. These findings show that based on capability, garlic extracts could provide benefit anti-infective therapy. On the other side, CHX mouthrinse is effective in controlling plaque and gingivitis but can cause color changes in teeth. [17] In addition, result of in vitro study done by Arabaci et al. showed that CHX also has a few genotoxic and cytotoxic effects on human lymphocytes. [18]


  Conclusion Top


Both the garlic extract and CHX mouthwash (0.12%) were effective against salivary microbial population however, CHX mouthwash showed higher efficacy than garlic extract against salivary microbial population. Mouthwash containing garlic can be a promising alternative for the patients with lesser microbial counts. Garlic has antimicrobial activity with no side effects such as dental staining and other cytototoxic effects when compared to CHX mouthwash. Further clinical investigations are needed for standardization and preparation of garlic containing toothpastes and mouthwashes for the prevention of oral microbial diseases.

 
  References Top

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Amruthesh S. Dentistry and ayurveda - IV: Classification and management of common oral diseases. Indian J Dent Res 2008;19:52-61.  Back to cited text no. 1
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Cowan MM. Plant products as antimicrobial agents. Clin Microbiol Rev 1999;12:564-82.  Back to cited text no. 2
    
3.
Saini R, Sharma S, Saini S. Ayurveda and herbs in dental health. Ayu 2011;32:285-6.  Back to cited text no. 3
[PUBMED]  Medknow Journal  
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Tsao SM, Yin MC. In-vitro antimicrobial activity of four diallyl sulphides occurring naturally in garlic and Chinese leek oils. J Med Microbiol 2001;50:646-9.  Back to cited text no. 4
    
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Fani MM, Kohanteb J, Dayaghi M. Inhibitory activity of garlic (Allium sativum) extract on multidrug-resistant Streptococcus mutans. J Indian Soc Pedod Prev Dent 2007;25:164-8.  Back to cited text no. 5
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Najafi MH, Taheri M, Mokhtari MR, Forouzanfar A, Farazi F, Mirzaee M, et al. Comparative study of 0.2% and 0.12% digluconate chlorhexidine mouth rinses on the level of dental staining and gingival indices. Dent Res J (Isfahan) 2012;9:305-8.  Back to cited text no. 6
    
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Shetty S, Thomas B, Shetty V, Bhandary R, Shetty RM. An in-vitro evaluation of the efficacy of garlic extract as an antimicrobial agent on periodontal pathogens: A microbiological study. Ayu 2013;34:445-51.  Back to cited text no. 7
[PUBMED]  Medknow Journal  
8.
Borhan-Mojabi K, Sharifi M, Karagah T. Efficacy of different concentrations of garlic extract in reduction of oral salivary microorganisms. Arch Iran Med 2012;15:99-101.  Back to cited text no. 8
    
9.
Khare CP. Text Book of Indian Herbal Remedies. Publisher Springer Berlin Heidelberg, 2012.  Back to cited text no. 9
    
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Bakri IM, Douglas CW. Inhibitory effect of garlic extract on oral bacteria. Arch Oral Biol 2005;50:645-51.  Back to cited text no. 10
    
11.
Kapoor LD. Handbook of Ayuvedic Medicinal Plants. CRC Press, 2000.  Back to cited text no. 11
    
12.
Sivam GP. Protection against Helicobacter pylori and other bacterial infections by garlic. J Nutr 2001;131:1106S-8.  Back to cited text no. 12
    
13.
Groppo FC, Ramacciato JC, Motta RH, Ferraresi PM, Sartoratto A. Antimicrobial activity of garlic against oral streptococci. Int J Dent Hyg 2007;5:109-15.  Back to cited text no. 13
    
14.
Yamada Y, Azuma K. Evaluation of the in vitro antifungal activity of allicin. Antimicrob Agents Chemother 1977;11:743-9.  Back to cited text no. 14
    
15.
Bachrach G, Jamil A, Naor R, Tal G, Ludmer Z, Steinberg D. Antimicrobial activity of garlic allicin on oral pathogen associated with dental caries and periodontitis. J Med Food 2011;14:1338-43.  Back to cited text no. 15
    
16.
Elnima EI, Ahmed SA, Mekkawi AG, Mossa JS. The antimicrobial activity of garlic and onion extracts. Pharmazie 1983;38:747-8.  Back to cited text no. 16
    
17.
Moreira AD, Mattos CT, de Araújo MV, Ruellas AC, Sant'anna EF. Chromatic analysis of teeth exposed to different mouthrinses. J Dent 2013;41 Suppl 5:e24-7.  Back to cited text no. 17
    
18.
Arabaci T, Türkez H, Çanakçi CF, Özgöz M. Assessment of cytogenetic and cytotoxic effects of chlorhexidine digluconate on cultured human lymphocytes. Acta Odontol Scand 2013;71:1255-60.  Back to cited text no. 18
    


    Figures

  [Figure 1], [Figure 2]
 
 
    Tables

  [Table 1], [Table 2]



 

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