Increase family Rutaceae. It has a distinct aroma

Increase in multi drug resistant
bacteria had led to the need to search for new antimicrobials has increased.
Currently there has been increase in natural and herbal medicine because of its
lesser side effects and reduced toxicity.1 Curry leaves scientifically known as Murraya koenigii are a popular
leaf-spice. It is a tropical to sub-tropical tree in the family Rutaceae. It has a distinct aroma due to the
presence of volatile oil. The leaves have a slightly pungent, bitter in taste,
and acidic in nature. Curry leaf is also used in many traditional cultures
namely Indian, Ayurvedic and Unani prescriptions.2 The
curry leaves contain proteins, carbohydrate, fiber, minerals, carotene,
nicotinic acid, Vitamin C, Vitamin A, calcium and oxalic acid. It also contains
crystalline glycosides, carbazole alkaloids, koenigin, girinimbin,
iso-mahanimbin, koenine, koenidine and koenimbine, Triterpenoid alkaloids cyclomahanimbine,
tetrahydromahanimbine. Murrayastine, murrayaline, pyrayafoline carbazole
alkaloids and many other chemicals are present.3 They
contain several medicinal properties such as antidiabetic, antioxidant, antimicrobial,
anti-fungal, anti-inflammatory, anti- carcinogenic and hepato-protective properties.

Bacteria are the etiological agents of periodontal
diseases, which remain the primary cause of tooth loss in adults. Periodontal diseases are usually initiated by plaque biofilm formation
which in turn gets mineralized into calculus. This plaque biofilm and calculus
act as a resident for many pathologic and physiologic bacteria including Porphyromonas
gingivalis, Staphylococcus aureus, Streptococcus sanguis, Streptococcus oralis
Treponema denticoli etc. Many researches have proved the presence of Staphylococcus
aureus in calculus.

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Staphylococcus aureus is a gram-positive, round-shaped bacterium that is a member of the Firmicutes, and it is a member of the normal flora of the body,
frequently found in the nose, respiratory tract, and on the skin. It is a facultative anaerobe. Staphylococcus aureus produces various
enzymes such as coagulase which clots plasma and coats the bacterial cell,
probably to prevent phagocytosis. Hyaluronidase breaks down hyaluronic acid and helps in spreading it. Staphylococcus aureus also produces deoxyribonuclease, which breaks down the DNA, lipase to digest lipids, staphylokinase to dissolve fibrin and aid in spread, and beta-lactamase for drug resistance. Antibiotic resistant strain
of Staphylococcus aureus is methicillin resistant Staphylococcus aureus4
which has become a clinical problem worldwide. It is important to bring out new
antibiotics against staphylococcus species.

Chlorhexidine is a biguanide compound
used as an antiseptic agent with topical antibacterial activity. Chlorhexidine is positively charged and reacts with
the negatively charged microbial cell surface, thereby destroying the integrity
of the cell membrane. Subsequently, chlorhexidine penetrates into the cell and causes leakage of
intracellular components leading to cell death. Since gram positive bacteria
are more negatively charged, they are more sensitive to this agent.

 

The aim of the study is to know the antibiotic effect
of curry leaves on Staphylococcus aureus.

 

MATERIALS AND METHOD:

 

Extract preparation:

Fresh curry leaves are collected and cleaned properly
with water. Leaves are dried completely and powdered. 30g of powder is
collected. 15 g of powder is mixed with 50 ml of water and other 15 g to
ethanol. The mixture is kept for 24 hrs with periodic shaking then filtered
and collected.

Culture Media:

Nutrient agar
was used as culture media. 14 gms of nutrient agar is mixed in 500 ml of
distiller water and heated till the agar is completely dissolved. The mixture
is autoclaved for 15 minutes for 121.
After autoclaving the mixture is poured in culture plates using micropipette
and the mixture is allowed to set.

Microbial Assay:

A subculture for
staphylococcus aureus was made by rubbing the swab containing staphylococcus
aureus strain over nutrient agar. The culture plate is left for 24 hours. After
sub culture the study contained 2 groups. Group 1 consist of curry leaves
extract with water and group 2 consist of curry leaves extract with ethanol.

Zone of
inhibition test- Agar well diffusion test was used to check the antimicrobial
activity of curry leaves against staphylococcus aureus. In each culture plate 3
wells were made and each well was filled with saline, chlorohexidine mouth wash
and last one with extract at 50  concentration. These culture plates were
incubated for 24 hours in hot air oven at 37.
Once the zone of inhibition is formed it is measured using ruler in
millimeters.

A-   
Saline

B-   
Chlorohexidine

C-   
Ethanolic extract

ZONE
OF INHIBITION WITH ETHANOLIC EXTRACT

 

 

A-   
Saline

B-   
Chlorohexidine

C-   
Aqueous extract

 

ZONE
OF INHIBITION WITH AQUEOUS EXTRACT

 

 

 

RESULT:

The
antimicrobial activity of the curry leaves extract at 50  concentration was screened by agar well
diffusion method and the zone of inhibition was measured in millimeters. Plate
1 – with water extract showed no changes except of chorohexidine with zone of
inhibition of 26mm whereas in plate 2- with ethanolic extract shows zone of
inhibition of 25 mm and chlorohexidine with 24 mm of zone of inhibition.

 

DISCUSSION:

The
antimicrobial activity of the curry leaves extract at 50  concentration was screened by agar well
diffusion method and the zone of inhibition was measured in millimeters. Plate
1 – with water extract showed no changes except of chorohexidine with zone of
inhibition of 26mm whereas in plate 2- with ethanolic extract shows zone of
inhibition of 25 mm and chlorohexidine with 24 mm of zone of inhibition.

Many
research proved the presence of staphylococcus aureus in sub – gingival and supra
– gingival calculus. Ohara-Nemoto et al.5
found numbers of staphylococci ranging from 102 to 105 CFU
g-1, albeit from supra-gingival plaque.  In this study they collected saliva and supra-gingival
calculus and isolated 9 Staphylococcus species and 334 isolates were
identified. Heller et al.6 analyzed the prevalence and
infection levels of 51 microbial species in the subgingival biofilm of 260
patients with chronic or aggressive periodontitis. They observed that Staphylococcus aureus was more prevalent in the
subgingival biofilm of patients with chronic periodontitis. In this study they
analysed that chronic periodonditis patient are usually individual leass than
or equal to 35 age of years and are non-smokers. Fritschi BZ et al.7 found
a strong association between Staphylococcus
aureus and aggressive periodontitis in
non-smokers. Although they hypothesized that the subgingival microbiota
does not differ between sites in individuals with chronic or aggressive
periodontitis, or by smoking status. Murdoch
et al.8 who isolated
staphylococci from at least one diseased site in 54% of periodontal patients,
and Loberto et al.9 who isolated staphylococci from the subgingival
biofilm in 37.5% of subjects with chronic periodontitis. Rams et al.10 isolated staphylococci
from 50.4% of patients with advanced adult periodontitis, and Dahlén and
Wikström 11 isolated Staphylococcus from 54.4% of patients. Few studies have found staphylococci in subgingival
biofilms samples, but they are impossible to show if staphylococcus has
significant role in periodontal disease or not.12,13  

Tulika
Pandit et al.14 proved that methanolic extract>> ethanolic
extract>aqueous extract of papaya and curry leaves showed antimicrobial
activity against the tested organism. Prathyusha Akula et al.15 proved
the anti-bacterial effect of Murraya Koenigii against staphylococcus aureus
followed by Proteus vulgaris and Enterobacter aerogens. Manish Vats et al. 16
proved the antimicrobial effect of Murraya Koenigii against
staphylococcus aureus, Micrococcus luteus, Bacillus subtilis, Escherichia coli,
Pseudomonas aeruginosa, Candida albicans and Aspergillus niger.  Ito et al,17 stated
that antimicrobial activity of roots of M. koenigii is due to presence
of carbazole alkaloids.

 

 

 

 

 

CONCLUSION:

Plants
are believed to have potential therapeutic effect. The secondary
metabolites of plants were found to be source of various phytochemicals that
could be directly used as intermediates for the production of new drugs.  It is important to bring about the use of
herbs in dentistry to decrease the side effects of synthetic medicine.
Ethanolic extract of curry leaves expressed antimicrobial activity at 50 concentration.
But the exact mechanism behind is still unknown.