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THE ANTIBACTERIAL EFFECT OF CURRY LEAVES (Murraya Koenigii)

Article in European Journal Pharmaceutical and Medical Research · October 2016

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 ejpmr, 2016,3(10), 382-387. SJIF Impact Factor 3.628
Research Article
Irfan et al. EUROPEAN JOURNAL OF PHARMACEUTICAL
European Journal of Pharmaceutical and Medical Research
AND MEDICAL RESEARCH ISSN 2394-3211
www.ejpmr.com EJPMR

THE ANTIBACTERIAL EFFECT OF CURRY LEAVES (Murraya Koenigii)

Hanan Al Harbi1, Dr. Uma M. Irfan2* and Dr. Sarah Ali3

1
Department of Medical Laboratories, College of Applied Medical Sciences, Qassim University, Buraydah, Al Qassim,
Saudi Arabia.
2
Professor of Epidemiology, Department of Medical Laboratories, College of Applied Medical Sciences, Qassim
University, Buraydah, Al Qassim, Saudi Arabia.
3
Assistant Professor, Microbiology, Department of Medical Laboratories, College of Applied Medical Sciences,
Qassim University, Buraydah, Al Qassim, Saudi Arabia.

Corresponding Author: Dr. Uma M. Irfan

Professor of Epidemiology, Department of Medical Laboratories, College of Applied Medical Sciences, Qassim University, Buraydah, Al
Qassim, Saudi Arabia.

Article Received on 22/08/2016 Article Revised on 12/09/2016 Article Accepted on 01/10/2016

ABSTRACT
Introduction: Curry Leaf (Murraya koenigii) is a green leafy vegetable native to India. Curry leaves are natural
flavoring agents with numerous health benefits. They contain several medicinal properties that include it being
anti-diabetic, antioxidant, antimicrobial, anti-inflammatory, anti-carcinogenic and with hepato-protective
properties. The aim of this study was to investigate the antibacterial effects of curry leaves. Methods: The curry
leaves samples were obtained from India. The leaves were dried at room temperature and pulverized to make curry
leaf powder. This powder was used to prepare the ethanol and methanol extracts to test the antimicrobial activity
on gram positive and gram negative bacteria; by the paper disc diffusion method. The inhibition zones around the
paper disc was measured and compared with standard measurements using antibiotic samples. Result: The curry
leaf extract showed a broad spectrum of very significant antibacterial activity by producing a clear zone of
inhibition against, Staphylococcus, E.coli, Streptococcus and Proteus. Both the ethanol and methanol extracts of
curry leaves were found to be effective against tested bacterial strains except, Klebsiella pneumonia and
Pseudomonas aeruginosa. The clear zones of inhibition in bacterial samples produced by curry leaf extracts were
comparable with antibiotics such as Gentamycin and Amikacin. Conclusion: Murraya koenigii extracts have
demonstrated antibacterial effects particularly on E.coli and Staphylococcus as compared to antibiotics such as
Gentamycin and Amikacin. Thus, curry leaves could be effective for prevention of bacterial infections and may be
considered as an alternative to antibiotic regimens.

KEYWORDS: Murraya koenigii, Bacteria, Inhibition zone, Antimicrobial effects.

INTRODUCTION Ayurvedic and Unani prescriptions (Suman Singh et al,

Indiscriminate use of antibiotics in today’s world is one 2014).
of the major causes for the rising emergence of multi
drug resistant pathogenic strains that do not respond to The leaves of Murraya koenigii contain proteins,
the usual line of treatment. Therefore, the need to search carbohydrate, fiber, minerals, carotene, nicotinic acid,
for new antimicrobials remains unchallenged. Currently, Vitamin C, Vitamin A, calcium and oxalic acid. It also
in addition to antibiotics and chemically synthesized contains crystalline glycosides, carbazole alkaloids,
drugs, the trend to look out for alternative medicines koenigin, girinimbin, iso-mahanimbin, koenine,
such as natural or herbal medicines is increasing because koenidine and koenimbine. Triterpenoid alkaloids
they may have fewer side effects or toxicity owing to cyclomahanimbine, tetrahydromahanimbine are also
their natural sources (Anita Joshi et al, 2009). present in the leaves. Murrayastine, murrayaline,
pyrayafoline carbazole alkaloids and many other
Curry leaves (Murraya koenigii) are a popular leaf-spice chemicals have been isolated from Murraya koenigii
used in very small quantities for their distinct aroma due leaves (Bhandari PR., 2012).
to the presence of volatile oil and their ability to improve
digestion. These leaves are widely used in Asian cuisines BACKGROUND
for flavoring foods. The leaves have a slightly pungent, Curry Leaf (Murraya koenigii) is an important leafy
bitter and feebly acidic taste, and they retain their flavor vegetable that belongs to the Rutaceae family, and is
and other qualities even after drying. Curry leaf is also native to India and the Southeast Asian Region. Curry
used in many traditional cultures namely Indian leaves are natural flavoring agents with a number of

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Irfan et al. European Journal of Pharmaceutical and Medical Research

important health benefits. They contain several medicinal This study was undertaken to evaluate the antibacterial
properties such as anti-diabetic, antioxidant, properties of the ethanolic and methanolic extracts of
antimicrobial, anti-fungal, anti-inflammatory, anti- curry leaves (Murraya Koenigii) on selected clinically
carcinogenic and hepato-protective properties. The pathogenic bacterial isolates.
various notable pharmacological activities of the plant
include activity on heart, anti-diabetic and cholesterol METHODOLOGY
reducing property, antimicrobial activity, antiulcer The plants used in this study were obtained from
activity, antioxidative property, cytotoxic activity, Southern India, where it is cultivated and exported to
antidiarrheal activity, phagocytic activity. The chemical Saudi Arabia. The curry leaves were shade dried at room
composition of the fresh leaves of Murraya koenigii temperature for 3 days and powdered using an electric
consists of volatile oil. Carbazole alkaloids and triterpene blender.
have been isolated from stem bark and roots of Murraya
koenigii (Bhandari PR, 2012 and Disegha GC., et al Methanol extraction: The methanol extract was made
2014). by, adding ten grams (10gms) of curry powder to 100 ml
of 70% aqueous methanol solution (w/v) covered with
Ethanol extracts of A. nilagirica (IC50 300 μg/ml) and filter paper kept on rotary shaker for 24 hours, and then
M. koenigii (IC50 400 μg/ml) were found to be more kept in a dark area at room temperature for 2 to 3 days.
effective against Mycobacterium smegmatis as compared The filtered supernatant was collected and the solvent
to petroleum ether and water extracts. M. koenigii extract was evaporated to make the final volume of the curry
showed maximum activity against M. bovis BCG in leaf methanol extract for the experiment.
combination with a first line anti-TB drug rifampicin. M.
koenigii leaf extract also exerted more cytotoxic (IC50 Ethanol extraction: The ethanol extract was made by,
20 μg/ml), genotoxic and apoptosis in mouse dissolving ten grams (10gms) of curry leaves powder in
macrophage RAW 264.7 cell line. Treatment of mouse 100 ml of ethanol and distilled water (8: 2 w/v) covered
macrophages with A. nilagirica extract increased with filter paper kept for 24 hours in a dark area at room
intracellular killing of M. smegmatis by inducing temperature. The filtered supernatant was collected and
production of reactive oxygen species and nitric oxide the solvent was evaporated by incubating at room
(Naik SK et al, 2014 and Elliot T. et al, 2011). temperature for 48 hours, to make the final volume of the
curry leaf ethanol extract for the experiment.
The leaf extract exhibited the highest antibacterial
activity against E. coli with 15±3mm zone of inhibition Determination of antimicrobial activity: Antibacterial
and for fungi the leaf extract exhibited more anti-fungal activity of the ethanolic and methanolic extracts of the
activity as in Aspergillus niger with a 14±1mm zone of curry leaves sample was evaluated by the paper disc
inhibition (Disegha GC et al 2014). The administration diffusion method. The disc made from filter paper
of aqueous extract of both the curry and olive leaves has (Whatman No3) with 6mm diameter, was soaked in
shown a significant decrease in the blood glucose level in methanolic and ethanolic solution (15 µl microliter in
STZ-induced diabetic rats (Maha El Amin et al, 2013). each disc) with the following concentrations: 100mg/ml;
50mg/ml; 25mg/ml; 12.5mg/ml and 6.25mg/ml; of
In a study by Rajendran MP et al (2014), the chemical leaves extraction of each extract (Collee JG, et al 1996).
constituents of the essential oil obtained from curry
leaves, identified using Gas Chromatography-Mass Disc diffusion tests: A bacterial suspension in sterile
Spectroscopy (GCMS) exhibited potent inhibition normal saline was prepared with reference to the 0.5
against antibiotic resistant bacteria such as McFarland Standards. Turbidity of the bacterial
Staphylococcus aureus; Pseudomonas aeruginosa; suspension was compared with 0.5 McFarland standard
Klebsiella pneumonia; Escherichia coli and solutions, followed by lawn culture of the bacterial
Streptococcus pyogenes. A significant minimum suspension on Mueller-Hinton agar plates using sterile
inhibition concentration values ranging from 25.0 to cotton swab and allowed to remain in incubator for 15
175.0 mg/ml and minimum bactericidal concentrations minutes. Then, the prepared methanolic and ethanolic
ranging from 100.0 to 500.0 mg/ml have been reported discs made from filter paper were placed on these
(Harika VC, et al 2010; Nagappan T, et al 2011; and cultures and the plates incubated at 37ºc for 24 hours.
Kamaraj C, et al 2014). The inhibition zone around each disc was measured in
millimeters (mm) and the assay was carried out three
The results of another study (Mohar Singh A et al, 2011), times for each extract to obtain consecutive results. The
revealed that most of the bacterial strains (except E. coli, four standard antibiotics discs used to compare the curry
B. cereus and S. faecalis) exhibited intermediate effect at leaves microbial inhibition activity were Amikacin,
low concentrations of leaf extract (at 10% and 15%) of Ciprofloxacin, Amoxicillin, and Gentamicin.
Murraya koenigii. However, those authors stated that the
efficacy of the leaf extract could have been increased by Antimicrobial-containing filter paper discs were placed
increasing the concentration of the extract. on agar plates inoculated with the bacteria to be tested
and incubated. Subsequently, fully susceptible bacteria

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Irfan et al. European Journal of Pharmaceutical and Medical Research

exhibited a large zone of growth inhibition around the 12mm inhibition zone for E.coli. The 100mg/ml
disc; less sensitive (intermediate sensitive) isolates concentration produced the maximum inhibition zones
exhibited a smaller inhibition zone and resistant bacteria for both bacteria which progressively reduced with
indicated no clear zones or grew up to the disc edge. In further dilutions from 50mg/ml to 6.25mg/ml.
this study we used six isolates of bacterial species;
Staphylococcus aureus and Streptococcus sp. Figure 2 to be placed here.
representing gram positive bacteria; and Escherichia coli, In comparing the inhibition zones of Staphylococcus,
Klebsiella pneumonia, Proteus sp. and Pseudomonas Streptococcus, E.coli and Proteus in Muller Hinton agar,
aeruginosa representing gram negative bacteria (Mohar the curry leaves (methanol) extract was more effective
Singh A et al, 2011). with Staphylococcus in producing a maximum of 16mm
inhibition zone as compared to a maximum of 12.33 mm
Statistical Analysis: Each experiment was repeated in inhibition zone for Streptococcus. The 100 mg/ml
triplicate sets and the mean values for the inhibition dilution produced the maximum inhibition zones for both
zones from the triplicate results were employed to bacteria which progressively reduced with the dilutions
represent the antibacterial activity of the curry leaf from 50mg/ml to 6.25mg/ml.
extracts. Results were recorded as S (sensitive), I
(intermediate sensitive) or R (resistant). The results of Table 1 describes the inhibition zones produced by
sensitivity tests were expressed as (0) = (R) for no known antibiotics such as Gentamicin, Amikacin,
sensitivity, + for (below 6 mm) = (R) for low sensitivity, Amoxicillin, and Ciprofloxacin on several bacterial
++ (7- 12mm) = (I) for moderate sensitivity and +++ species namely: Staphylococcus Aureus, Streptococcus,
(13mm & above) = (S) for high sensitivity (Rajendran E.coli, Proteus, Klebsiella and Pseudomonas. In
MP, et al 2014). comparison the table also contains values on the
inhibition zones created by the curry leaf ethanol extract
RESULTS on the various bacterial species. It was evident that the
The antibacterial effects of curry leaves (Murraya curry leaf ethanol extract has produced a larger inhibition
Koenigii) extracts (with ethanol and methanol) as tested zone than all the four antibiotics particularly for
on bacterial samples and compared with known Staphylococcus bacteria, with the inhibition zone is 15
antibiotics are described with inhibition zones measured mm. The curry leaf ethanol extract did not have any
in millimeters. effect on bacterial species such as Proteus, Klebsiella and
Pseudomonas.
Figure 1 to be placed here.
In comparing the inhibition zones of Staphylococcus, The curry leaf methanol extract (Table 2) produced a
Streptococcus and E.coli on Muller Hinton agar, the larger inhibition zone than all the four antibiotics
curry leaves (ethanol) extract was more effective with particularly for Staphylococcus bacteria, with an
Staphylococcus in producing a maximum of 15mm inhibition zone of 16 mm. The curry leaf methanol
inhibition zone as compared to a maximum of 13 mm extract did not have any effect on bacterial species such
inhibition zone for Streptococcus and to a maximum of as Klebsiella and Pseudomonas.

Figure 1: Comparison of inhibition zone for various dilutions of ethanol extraction of curry leaves and effect on
Staphylococcus sp., Streptococcus sp., and E.coli.

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 Irfan et al. European Journal of Pharmaceutical and Medical Research

Figure 2: Comparison of inhibition zone for various dilutions of Methanol extraction of curry leaves and effect
on Staphylococcus sp., Streptococcus sp., E.coli, and Proteus sp.

Table 1: Distribution of the Inhibition Zones (in millimeters) produced by Murraya Koenigii (ethanol
extraction) on bacteria in Muller Hinton agar compared with antibiotics: Amikacin, Ciprofloxacin, Amoxicillin
and Gentamicin.
Ethanol Ethanol Ethanol Ethanol Ethanol
Bacterial species GN AK AX CIP
100 dilution 50 dilution 25 dilution 12.5 dilution 6.25 dilution
Staphylo-coccus 11.00 7.00 8.00 9.00 15.00+++ 13.33+++ 12.33++ 8.33++ 2.33+
Strepto-coccus 13.00 11.00 8.00 6.00 13.00+++ 12.00++ 10.67++ 8.67++ 2.67+
E.coli 12.00 14.00 12.00 15.00 12.00++ 10.33++ 7.33++ 6.33+ 2.00+
Proteus 12.00 7.00 9.00 9.5 0.00 0.00 0.00 0.00 0.00
Kleb-siella 12.00 8.00 8.00 9.00 0.00 0.00 0.00 0.00 0.00
Pseudo-monas 12.00 13.00 12.00 14.00 0.00 0.00 0.00 0.00 0.00
AK= amikacin; AX= amoxicillin; CIP=Ciprofloxacin; GN=Gentamicin
0 = No Sensitivity; + = Low Sensitivity; ++ = Moderate Sensitivity; +++ = High Sensitivity

Table 2: Distribution of the Inhibition Zones (in millimeters) produced by Murraya Koenigii (methanol
extraction) on bacteria in Muller Hinton agar compared with antibiotics: Amikacin, Ciprofloxacin, Amoxicillin
and Gentamicin.

Bacterial Methanol1 Methanol50 Methanol 25 Methanol12 Methanol6.2

GN AK AX CIP
species 00 dilution dilution dilution .5 dilution 5 dilution
Staphylo-coccus 11.00 7.00 8.00 9.00 16.00+++ 15.67+++ 14.00+++ 10.33++ 9.00++
Strepto-coccus 13..00 11.00 8.00 6.00 12.33++ 11.00++ 9.33++ 7.67++ 2.33+
E.coli 12.00 14.00 12.00 15.00 4.33+ 3.33+ 3.33+ 3.00+ 2.67+
Proteus 12.00 7.00 9.00 9.5 3.33+ 3.00+ 3.00+ 2.67+ 0.000
Kleb-siella 12.00 8.00 8.00 9.00 0.00 0.00 0.00 0.00 0.00
Pseudo-monas 12.00 13.00 12.00 14.00 0.00 0.00 0.00 0.00 0.00
AK= amikacin; AX= amoxicillin; CIP=Ciprofloxacin; GN=Gentamicin
0 = No Sensitivity; + = Low Sensitivity; ++ = Moderate Sensitivity; +++ = High Sensitivity

DISCUSSION health of human beings (Anita Joshi et al, 2009) In this

Indiscriminate use of antibiotics has led to the rising context curry leaves have displayed immense potential as
emergence of multi drug resistant pathogenic strains of that natural alternative, especially with its antimicrobial
bacteria causing diseases that challenge regular treatment property.
protocols. Currently, in addition to antibiotics and
chemically synthesized drugs, the trend to look out for Curry leaf oil extracts have demonstrated (Rajendran MP
alternative medicine in nature is increasing as the natural et al, 2014) the strongest inhibition zone against Proteus
resources are less toxic and less deleterious to the overall mirabilis (18mm), Staphylococcus aureus,

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Irfan et al. European Journal of Pharmaceutical and Medical Research

Corynebacterium pseudotuberculosis (15mm), Klebsiella is evident that curry leaves are an effective natural
pneumoniae (15mm) Pseudomonas aeruginosa (14mm), remedy for bacterial infections that provide therapeutic
Enterobacter aerogenes (13 mm) and a moderate level effects with little or no side effects in combatting
zone of inhibition was observed with Salmonella enterica bacterial infections in humans.
(11mm), and Streptococcus pyrogens (10 mm)
respectively. Also, in this study Murraya Koenigii CONCLUSION
displayed antibacterial activity against the Gram-positive Murraya koenigii (Curry leaves) extracts have
and Gram-negative bacteria, while Escherichia coli demonstrated antibacterial effects particularly on E.coli,
showing maximum susceptibility to acetone extract with Staphylococcus, Streptococcus and Proteus as compared
a zone of inhibition of 16 mm. Our study had comparable to antibiotics such as Gentamycin and Amikacin in our
results with the inhibition zone for E.coli ranging from study. The ethanol and methanol extracts of curry leaves
12mm to 24mm by the various methods employed in the were found to be effective against all tested bacterial
experiment. strains except, Klebsiella pneumonia and Pseudomonas
aeruginosa. M. koenigii has the bioactive potential and it
Another study by Das and Biswas (2012) indicated that may be the best natural alternative to antibiotic therapy
ethyl acetate and dichloromethane soluble partition of for tested microbes. Therefore, curry leaves could be
methanolic leaf extract of Murraya Koenigii exhibited effectively used as a natural remedy in everyday meal,
mild activity against Escherichia coli forming zone of for the prevention of bacterial infections. Indeed this
inhibition of 9 mm to 11 mm. In our study the ethanol phenomenal plant may serve as a useful resource in the
extract of Murraya Koenigii exhibited an even larger food industry and clinical medicine. This research could
inhibition zone of 24 mm for E.coli followed by 12mm be further extended to test the bioactive properties of
to 15mm for Staphylococcus sp. However, the curry curry leaves for therapeutic use.
leaves methanol extract was more effective with
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