EVALUATION OF ANTIMICROBIAL EFFICACY OF PSIDIUM GUAJAVA, ACACIA ARABICA, PUNICA GRANATUM ON STREPTOCOCCUS MUTANS, LACTOBACILLI, PORPHYROMONAS GINGIVALIS, ENTEROCOCCUS FEACALIS AND CANDIDA ALBICANS.
BRIEF RESUME OF THE INTENDED STUDY:
6.1 Need for the study:
Dental biofilms constitute an ecosystem of microorganisms. The major microorganisms includes Streptococcus mutans, Lactobacilli,Porphyromonas gingivalis, Enterococcus feacalisand Candida albicans which are responsible for dental caries, endodontic and periodontal diseases. Controlling the levels of these microorganisms in the oral biofilm will aid in the prevention of these diseases of the oral cavity.
Numerous antimicrobial agents have been used to eliminate these microorganisms from the oral cavity of which chlorhexidine is the most commonly used mouth rinse and root canal irrigant. The incorporation of broad spectrum antimicrobial mouth rinses as adjuncts to oral hygiene regimens has assumed greater importance with the recognition that most individuals are unable to maintain adequate oral hygiene. Excessive use of these agents can result in microbial resistance, imbalance in oral flora and numerous adverse effects. These problems necessitate further research for alternate novel antimicrobial agents that are safe and potent against specific oral pathogens. Phytochemicals derived from medicinal plants have proven to have high antimicrobial, anti-inflammatory, anti-oxidant and biocompatible properties. Few amongs them are,
Punica granatum (pomegranate): This plant has been used to treat various ailments in traditional medicine. It has been noted for its antimicrobial and antioxidant potency.
Psidium guajava (guava): This tropical tree has been used as an effective and reliable aid in cleaning teeth in folk medicine. It also has anti-diarrhoeal, anti-inflammatory, anti-oxidant properties etc.
Acacia arabica (Indian gum arabica): The gum of this tree has been used for thousands of years for various clinical implications. This gum has the potential to inhibit early plaque formation and gingival inflammation.
The purpose of this study is to evaluate and compare the antimicrobial efficacy of guava leaves, pomegranate fruit juice and Acacia arabica gum on S. mutans, Lactobacillus, E. faecalis, P. gingivalis and C. albicans.
6.2 Review of literature:
In a study by Saraya S et al (2008), the anticariogenic activity of guava leaves extracts against S. mutans by agar diffusion method was studied. The results showed significant bactericidal activity against S. mutans.1
Pradeep AR et al (2010) compared Acacia arabica gel and chlorhexidine gel and evaluated their antiplaque effects. Patients on Acacia arabica showed significant clinical improvement in gingival and plaque scores compared to other patients.2
In a study conducted by Jebashree HS et al (2011) the plant P. guajava was initially evaluated for the anticariogenic activity against S. mutans by the disc diffusion method. The ethyl acetate extract of this plant showed high antibacterial activity against S. mutans. The antifungal activity was tested against C. albicans and the results revealed that P. guajava extract showed siginificant antifungal activity against C. albicans.3
Alsaimary IE (2012) isolated S mutans, E coli, S aureus, Klebsiella, Proteus and Pseudomonas from 60 samples of dental caries and evaluated the antimicrobial acivity of pomegranate juice against these organisms. The pomegranate juice was found to be effective against S. mutans in comparison to the other microorganisms included in there study.4
A study conducted by Kote et al (2011) using pomegranate juice on the dental plaque microorganisms S. mutans and Lactobacillus showed significant reduction in the level of dental plaque microorganisms after rinsing with pomegranate juice.5
6.3 Aim of the study:
To evaluate and compare the antimicrobial efficacy of guava leaves, pomegranate fruit juice and Acacia arabica gum on S. mutans, Lactobacillus, E. faecalis, P. gingivalis and C. albicans.
To evaluate the antimicrobial efficacy of P. granatum against S. mutans, Lactobacillus, E. faecalis, P. gingivalis and C. albicans.
To evaluate the antimicrobial efficacy of Acacia arabica against S. mutans, Lactobacillus, E. faecalis, P. gingivalis and C. albicans.
To evaluate the minimal inhibitory concentration of the extracts of P.gaujava, P.granatum, Acacia arabica on S. mutans, Lactobacillus, E. faecalis, P. gingivalis and C. albicans.
To compare the antimicrobial efficacy of P.guajava, P.granatum, Acacia arabica on S. mutans, Lactobacillus, E. faecalis, P. gingivalis and C. albicans.
MATERIALS AND METHODS
7.1 Source of data:
Source of samples: Gauva leaf powder and Acacia arabica gum will be obtained from the NKCA Pharmacy Ltd., Mysore. Commercially available fresh pomegranate will be obtained.
Standard ATCC microbial cultures S.mutans (ATCC25175), Lactobacillus fermentum (ATCC9338), P.gingivalis (ATCC33277), E.faecalis (ATCC29212), C.albicans (ATCC10231) will be procured from Mercury Instruments and Chemicals, Mysore.
Study set up: Department of Oral Pathology and Microbiology, Farooqia Dental College and hospital.
Study Design: An in-vitro study will be conducted to evaluate the antimicrobial efficacy of these medicinal plants on S.mutans, Lactobacillus,E.faecalis,P.gingivalis and C.albicans.
7.2 METHOD OF COLLECTION OF DATA:
Sample size- Each of the 3 plant extracts will be tested against each of the 5 microorganisms in sets of 6 cultures, giving a total number of 90 culture tests.
Guava leaves extract
Acacia Arabica gum extract
Pomegranate juice extract
0.2% Chlorhexidine solution
S. mutans, Lactobacillus, E. faecalis, P. gingivalis, C. albicans
Guava leaf extract and Acacia arabica gum extract will be prepared with methanol and distilled water as solvents. Pomegranate juice extract will be prepared from the procured pomegranates.
Microbial cultures will be maintained on brain heart infusion (BHI) broth and bacterial growth will be checked by changes in turbidity at 24 hrs by using the McFarland scale.
Well agar diffusion method will be performed to test the antimicrobial efficacy of the above extracts. BHI (brain heart infusion) agar and PDA (potato dextrose agar) plates will be prepared and cultures (200μl) will be inoculated by spreading on to agar plates by using sterile swab brush across the media. Four round wells, 4mm deep and 8mm diameter shall be punched using sterile cork borer and it will be marked as A, B, C, and D.
Group A – Gauva leaf extract
Group B – Acacia Arabica gum extract
Group C – Pomegranate juice
Group D – Chlorhexidine
Guava leaf extract, pomegranate juice, acacia arabica gum extract shall each be added to the respective wells and the bacterial plates will be incubated for 24hrs at 370C in an incubator. C.albican plates will be incubated at 280C for 72 hrs. After the incubation period, the plates will be removed and zones of inhibition will be measured in millimeters using vernier calliper and statistically analysed.
The minimum inhibitory concentration (MIC) will be determined for all the 3 plant extracts against S. mutans, Lactobacillus, E. faecalis, P. gingivalis and C. albicans as per the standard procedure followed by National Committee for Clinical Laboratory Standards(NCCL).
Duration of study: 6 months - 1 year.
7.4 STATISTICAL ANALYSIS:
The following statistical procedures will be used to analyze the data.
1. Descriptive statistics
2. Chi- square test
3. Contingency coefficient analysis
4. T test
Statistical analysis will be done using ‘ Statistical Package for Social Sciences (SPSS)’ software using Windows V16.
7.5 Does the study require any investigations or interventions to be conducted on patients or other humans or animals? If so, please describe briefly.
7.6 Has ethical clearance been obtained from your institution in case of 7.5?
No LIST OF REFERENCES:
1. Saraya S, Kanta J, Sarisuta N et al. Development of guava extract chewable tablets for anticariogenic activity against Streptococcus mutans. J of Pharma Sci. 2008;35(1-4):18-23.
2. Pradeep AR, Agarwal E et al. Clinical and microbiologic effects of commercially available gel and powder containing Acacia arabica on gingivitis. Aust Dent J. 2010;57:312-8.
3. Jebashree HS, Kingsley SJ, Sathish ES, Devapriya D. Antimicrobial activity of few medicinal plants against clinically isolated human cariogenic pathogens- An In Vitro Study. International Scholarly
Research Network. 2011;1-6.
4. Alsaimary IE. Efficacy of some antibacterial agents against Streptococcus mutans associated with tooth decay. Afri J of Biotech. 2012;11(88):15457-9.
5. Kote S, Nagesh L. Effect .of pomegranate juice on dental plaque microorganisms (Streptococci and Lactobacilli). Anc Sci Life. 2011;31(2):49-51.