In the study1

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					        RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES, BANGALORE

                                 KARNATAKA

        PROFORMA FOR REGISTRATION OF SUBJECTS FOR DISSERTATION



                                    Dr.KESHAV SINGLA,
1        NAME OF THE CANDIDATE      DEPARTMENT OF CONSERVATIVE
            AND ADDRESS             DENTISTRYAND ENDODONTICS,
                                    DR.SYAMALA REDDY DENTAL
                                    COLLEGE, HOSPITAL&RESEARCH
                                    CENTRE,
                                    MUNNEKOLALA, MARATHAHALLI,
                                    BANGALORE-37

                                    DR.SYAMALA REDDY DENTAL
    2    NAME OF THE INSTITUTION    COLLEGE, HOSPITAL & RESEARCH
                                    CENTRE, MUNNEKOLALA,
                                    MARATHAHALLI,
                                    BANGALORE – 560037.



                                    MASTER OF DENTAL SURGERY
    3     COURSE OF STUDY AND       CONSERVATIVE DENTISTRY &
             SUBJECT                ENDODONTICS



    4     DATE OF ADMISSION TO           01 JUN – 2011
             COURSE



    5   TITLE OF THE TOPIC

        “ COMPARISON OF THE ANTIBACTERIAL EFFICIENCY OF NEEM
        LEAF,TULSI LEAF EXTRACTS AND TEA TREE OIL WITH 2%
        SODIUM HYPOCHLORITE AGAINST E.FAECALIS- AN IN VITRO
        STUDY”
6   BRIEF RESUME OF INTENDED WORK:

    6.1 NEED FOR THE STUDY:

           The major objective in root canal treatment is to disinfect the entire root
    canal system. Although cleaning, shaping, and use of antimicrobial medicaments are
    effective in reducing the bacterial load some bacteria do remain behind and
    multiply, causing reinfection of the canal.
           Considering the ineffectiveness, potential side effects and safety concerns of
    synthetic drugs, the herbal alternatives for endodontic usage might prove to be
    advantageous. NaOCl, chlorhexidine and calcium hydroxide have varying degree of
    antibacterial properties. But they have several undesirable characteristics such as
    tissue toxicity, risk of emphysema, allergic potential, disagreeable smell and taste,
    and inability to remove smear layer.
             Owing to the potential side effects, safety concerns and ineffectiveness of
    conventional allopathic formulations, consumption of preparations from Neem,
    Tulsi and Tea tree oil has increased over the last few decades. Herbs are used in
    dental and medical practice for long time. Now a days because of their high
    antimicrobial, anti-inflammatory, anti-oxidant and biocompatibility properties, their
    use in endodontics has become more popular.




    6.2 REVIEW OF LITERATURE:

    In the study1, the authors compared the antimicrobial efficacy of ginger extract and
    2% sodium hypochlorite against Enterococcus Faecalis using agar diffusion method.
    They concluded that ginger extract showed statistically significant activity against
    Enterococcus faecalis.
           In this in-vitro study2, the authors compared the antibacterial efficiency of
    neem leaf extract and 2% sodium hypochlorite against E.faecalis, C.albicans and
    mixed culture by using the agar diffusion test. They concluded that neem leaf extract
has a significant antimicrobial effect against E.faecalis, C.albicans and mixed
culture to use as intracanal medicament.
         In this study 3, the authors evaluated the antimicrobial activity of various
concentrations of Tulsi (Ocimum sanctum) extract against Streptococcus mutans by
agar diffusion test. They concluded that Tulsi extract demonstrated an antimicrobial
activity against Streptococcus mutans. It has maximum antimicrobial potential at the
concentration of 4% level.


        In this study4, the authors compared the inhibitory activity of root canal
irrigants against Candida albicans, Enterococcus faecalis and Staphylococcus aureus
by agar diffusion test and redox indicator rasazurin to check the inhibitory effect and
metabolic activities of these microorganisms. They concluded that 5.25% sodium
hypochlorite is most effective against E.faecalis as compared to EDTA and citric
acid.


        In this in vitro study5 , the authors checked the effect of mango and      neem
extract on four microorganisms causing dental caries: Streptococcus mutans,
Streptococcus salivavius, Streptococcus mitis, and Streptococcus sanguis: by agar
diffusion test. They concluded that neem extract produced the maximum zone of
inhibition on streptococcus mutans at 50% concentration. neem even at 5%
concentration it shows zone of inhibition for all four bacteria. Mango extract at 50%
concentration showed maximum zone of inhibition on Streptococcus mitis.




6.3 OBJECTIVES OF THE STUDY:
      1 To check the antibacterial efficiency of neem leaf,tulsi leaf extract and tea
tree oil against Enterococcus.Faecalis.

      2 To compare the antibacterial efficiency of neem leaf, tulsi leaf extract and
tea tree oil with 2% Sodium hypochlorite.
7   MATERIALS AND METHODS:
    7.1 SOURCE OF DATA

          Neem leaf extract
          Tulsi extract
          Tea tree oil
          2% Sodium hypochlorite solution
          Normal saline
          Enterococcus faecalis stains ( ATCC 29212)
          Brain heart infusion (BHI) broth powder
          Brain heart infusion (BHI) agar plates
          Incubator
          Autoclave
          Sterilized swabs
          Vernier caliper
           Cork borer
          Burner
          Flask
          Glass micropippets
          McFarland scale




    7.2 METHOD OF COLLECTION OF DATA
           Neem extract,tulsi extract, tea tree oil and fresh stains of E.faecalis(ATCC
    29212) shall be used.
                    E.faecalis cultures shall be maintained on brain heart infusion (BHI)
    broth. Cultures shall grow overnight at 370C in BHI broth and bacterial growth
    shall be checked by changes in turbidity at 24hrs by using the McFarland scale.
                       To check the antimicrobial efficacy of neem leaf extract, tulsi leaf
    extract, tea tree oil and 2% NaOCl, agar diffusion method shall be performed. BHI
    (brain heart infusion) agar plates shall be prepared and cultures (200μl) will be
    spread on to agar plates. Inoculation should be performed by using sterile swab
    brush across the media. Four round wells,4mm deep and 8mm diameter shall be
    punched using sterile cork borer. Named as groups A,B,C,D.
                  Group A – Neem leaf extract,
                  Group B – Tulsi leaf extract,
                  Group C – Tea tree oil,
                  Group D – 2% Sodium hypochlorite.        .
                       Neem leaf extract, tulsi leaf extract, tea tree oil and 2% sodium
    hypochlorite (50μl) each shall be added to the respective wells and the plates will be
    incubated for 24hrs at 370C in an incubator.
                       After incubation period, plates will be removed and zones of
    inhibition will be recorded with vernier caliper. Experiment will be performed 3
    times and mean of the zone of inhibition will be recorded in mm.
    -                  The results will be tabulated and statistically analyzed by using
    analysis of variance (ANOVA).


    7.3 Does the study require any investigation or interventions to be conducted on
    patients or other humans or animals? If so, please describe briefly.
                                       No
    7.4. Has ethical clearance been obtained from your institution in case of 7.3?
                                       NA



8   LIST OF REFERENCES
       1. Gulve N M, Gulve N D. Comparison of antimicrobial efficacy of ginger
          extract and 2% sodium hypochlorite against enterococcous faecalis using
          agar diffusion method. JIDA 2010; 4 (10):347-349.

        2. Bohora A, Hegde V, Kokate S. Comparison of antibacterial efficiency of
           neem leaf extract and 2% sodium hypochlorite against E.faecalis, C.albicans
           and mixed culture- an in vitro study. Endodont 2010; 22: 10-14.

        3. Agarwal P, Nagesh L, Murlikrishnan. Evaluation of the antimicrobial
           activity of various concentrations of Tulsi(Ocimum sanctum) extract against
           Streptococcus mutans: an in vitro study. Indian J Dent Res2010; 21(3):357-
           359.

        4. Barcelos R, Gleiser R, Silva F. Inhibitory activity of root canal irrigants
           against Candida albicans, Enterococcus faecalis and Staphylococcus aureus.
           Braz oral res 2010; 24(4):406-412.
         5.    Prashant GM, Chandu GN, Murulikrishna KS, Shafiulla. The effect
              of mango and neem extract on four organisms causing dental caries:
              Streptococcus mutans, Streptococcus salivavius, Streptococcus mitis, and
              Streptococcus sanguis: An in vitro study. Indian J Dent Res 2007;18(4):
              148-151.




9             Signature of Candidate:               DR. KESHAV SINGLA



10            Remarks of the guide               This is a genuine study, which will be
                                                 carried out by the post graduate student
                                                 under my supervision and guidance.


 11    NAME AND DESIGNATION OF



                                                         DR. UDAY KAMATH

      11.1                Guide                        PROFESSOR & HEAD
                                                    Dept of Conservative Dentistry &
                                                              Endodontics


      11.2              Signature



                                                           DR. HINA SHETH
      11.3           Co-guide (if any)
                                                             PROFESSOR
                                                    Dept of Conservative Dentistry &
                                                              Endodontics



      11.4               Signature
   11.5     Head of Department      DR.UDAY KAMATH




   11.6        Signature



12 12.1   Remarks of the Chairman
          and Principal




   12.2        Signature

				
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