Antimicrobial Activity of Commiphora myrrha Against Some Bacteria and Candida albicans Isolated from Gazelles at King Khalid Wildlife Research Centre
to cite this article:
S.A. Omer, S.E.I. Adam and O.B. Mohammed, 2011. Antimicrobial Activity of Commiphora myrrha Against Some Bacteria and Candida albicans Isolated from Gazelles at King Khalid Wildlife Research Centre. Research Journal of Medicinal Plants, 5: 65-71.
Received: April 02, 2010;
Accepted: June 15, 2010;
Published: July 14, 2010
Plant extracts have been used for a wide variety of purposes for thousands
of years (Jones, 1996). Commiphora myrrha is
a member of family Burseraceae, known locally as morr Hijazi or myrrh
and commercially as Arabian myrrh or Karam. Myrrh is the dried gum-resin from
a number of closely related, small, thorny trees of the genus Commiphora,
probably originating in the highlands of Yemen (Wadi Hadramaut). It is widely
distributed in the Kingdom of Saudi Arabia and it is grown in Jizan area on
Red Sea coast, a distinct so bare and dry that is called Tihama meaning very
hot hill. It is also found in Somalia and other coast African countries (Mugahid,
1981; Vollensen, 1985). It is used in traditional
medicine as antiseptic, carminative, anti-inflammatory, tonic in dyspepsia and
emmenogogue (Tarig et al., 1985). It also used
as remedy for spongy gums, aphthous stomatitis and indolent ulcer (Satyavati
et al., 1969). The Southern most countries on the Arabian Peninsula,
today Yemen and Oman, have exported myrrh (and olibanum) for incense ceremonies
in the temples and principalities of the Orient.
The constituents of this plant are terpenes, sesquiterpenes, aldyhydes, euginol,
resin commophoric acids, volatile and essential oils, salts and proteins (Provan
and Waterman, 1988; Al-Harbi et al., 1994).
The antimicrobial activity of plant oils and extracts has formed the basis of
many applications, including raw and processed food preservation, pharmaceuticals,
alternative medicine and natural therapies (Reynolds, 1996;
Lis-Balchin and Deans, 1997). Hammer
et al. (1999) studied the antimicrobial activity of essential oils
and extracts of 52 plants. Mothana and Lindequist (2005)
and Mothana et al. (2009) studied the antimicrobial,
anticancer and antioxidant activities of extracts from several plant species
from the Island Soqotra, among which they evaluated the antimicrobial and antifungal
activity of Commiphora parvifolia and C. ornifolia. On a different
study Abbas et al. (2007) evaluated the extracts
of C. opobalsamum as antitumour, antimicrobial, anti-inflammatory, antioxidant
and antimalarial as well as its estrogenic activity. The effect of C. mukul
extracts on the cardiac function was evaluated by Ojha et
al. (2008) and they reported significant improvement of cardiac function
and prevention of myocardial ischemic impairment was associated with the use
of this plant extracts.
In this study the effect of ethanolic and ether extracts of C. myrrha as antibacterial against some Gram-positive and Gram-negative organisms and as antifungal against Candida albicans isolated from gazelles at King Khalid Wildlife Research Centre is evaluated.
MATERIALS AND METHODS
One kilogram of C. myrrha oleo-gum resin was finely ground and successively
extracted with petroleum ether (60-80°C) for 18 h and with ethanol for 14
h using sexhlet apparatus. The oleo-gum resin of Commiphora myrrha extraction
was performed at the Department of Veterinary Medicine, Pharmacology and Toxicology,
University of Khartoum during 2008. The method used to evaluate antimicrobial
activity of the plant extracts was the two-layer technique described by Stokes
and Waterworth (1972). The media used was Muller Hinton agar medium (Laboratories
Britania S.A. Lopotos, Buenos Aires, Argentine). The composition of Muller Hinton
agar is 300 g L-1 beef extract, 175 g L-1 acid casein
hydrolase, 1.5 g L-1 starch and 15 g L-1 agar. Bacteriological
evaluation was done at the Veterinary Research Laboratory at King Khalid Wildlife
Research Centre, Saudi Arabia.
Thirty seven grams of the medium were dissolved in one liter of warm distilled water and the reconstituted medium was sterilized by autoclaving at 121°C for 15 min. The medium was then distributed in sterile Petri dishes (95 mm in diameter) in 10 mL volumes and allowed to cool forming the base layer. Two Gram-positive organisms, Bacillus subtilis and Staphylococcus albus and two Gram- negative organisms Escherichia coli and Pseudomonas aeruginosa were grown in nutrient broth (Oxoid) at 37°C for 10 h. Candida albicans was grown in Sabouraud Dextrose Agar (SDA) at 37°C for 4-5 days. These organisms were isolated from gazelles raised at King Khalid Wildlife Research Centre, Thumamah. A fresh Mueller Hinton agar medium was prepared the same way as the base layer and was inoculated with the test organisms including C. albicans at 106 cells mL-1. The inoculated medium was distributed evenly in 10 mL volumes into the surface of the base layer. The plates were stored in the refrigerator at 4°C till use. On each inoculated plate, 5 cups (8 mm diameter) cut using sterile cork porer. Concentrations of 10, 20, 40, 80 and 100 mg mL-1 of the ethanolic and the ether extracts were made up and 200 μL of each concentration was placed in one of the cups in each plate by mean of sterile Pasteur pipette. The extracts were allowed to diffuse for 3 h before incubating the plates for 18 h at 37°C. Oxytetercycline (Beecham, UK) was used as the reference drug at similar concentrations to those of extracts. Three replicates were made from each concentration. The diameter of inhibition zones resulting from the activity of the extracts were measured in mm and comparative activity was recorded.
RESULTS AND DISCUSSION
The ethanol extract of C. myrrha exhibited anti-bacterial activity against E. coli, P. aeruginosa and S. albus (Fig. 1) but did not show activity against B. subtillis and C. albicans. The Minimum Inhibitory Concentration (MIC) of ethanolic extract against both E. coli and S. albus was found to be 40 mg mL-1, while that against P. aeruginosa was shown to be 20 mg mL-1. The ether extract, on the other hand, was highly active against Gram positive organisms (B. subtillus, S. albus) and C. albicans (Fig. 2) but did not show any activity against Gram negative organisms used in the present investigation (E. coli and P. aeruginosa). The minimum inhibitory concentration (MIC) of the ether extract against both S. albus and Candida abicans was found to be 10 mg mL-1 unlike that MIC shown for B. subtilis which was 40 mg mL-1. In both ethanolic and ether extract the activities of the extracts increased with the increase of the concentration of the extract.
The ethanolic and ether extracts activities against the bacteria and the yeast used in this study were found to show lower activity compared to the control antibiotic (oxytetracycline) used (Table 1). However, unlike the oxytetratcycline used the ether extract showed consistent activity against C. albicans.
The present investigation have shown that the petroleum ether extract exhibited activity against S. albus, B. subtilis and C. albicans with no activity against E. coli or P. aeruginosa. The ethanolic extract was highly active against E. coli and P. aerguinosa and S. albus. This might possibly explain the common use of C. myrrha for the treatment of infections in the mouth such as mouth ulcers, gingivitis, phyorrhea, as well as the catarrhal problems of pharyngitis and sinusitis.
||Activity of ethanolic extract of Commiphora myrrha oleo-gum
resin against Staphylococcus albus. The increase in the zone of inhibition
is proportional to the increase of the extract
||Activity of ether extract of Commiphora myrrha oleo-gum
resin against Candida albicans. The increase in the zone of inhibition
is proportional to the increase of the extract
|| Antimicrobial activities of ethanolic and ether extracts
of Commiphora myrrha, zone of inhibition is measured in mm
The high activity of the ether extract against Candida albicans coincides
with the routine use of C. myrrha in Ayurveda for oral and vaginal
hygiene, parasite treatment, antiseptic action and as a natural antibiotic (Nadkarni,
1992; Treadway, 1998). The use of ether extract
may also help in treating laryngitis, respiratory complaints, thrush and in
footbath for athletes feet because of its anti-fungal properties demonstrated
in the present investigation.
The method of antimicrobial activity assessment and choice of test organisms
may vary from one plant species to anther (Janssen et
al., 1987). Hammer et al. (1999) reported
antimicrobial activity of C. myrrha oil against many organisms using
two different methods. Unlike what we have demonstrated in the present investigation,
Hammer et al. (1999) found that oil from C.
myrrha inhibits Gram positive organisms only. Ethanolic and ether extracts
of C. myrrha used in this investigation showed remarkable activity against
Gram positive and Gram negative organisms as well as against C. albicans.
This can be attributed to changes in the composition of plant extract from different
geographical regions (probably different plant species), methods used for assessment
of the activity as well as the method used for extraction (Janssen
et al., 1987; Sivropoulou et al., 1995;
Reynolds, 1996). The oil preparation used by Hammer
et al. (1999) may contain certain extracts of C. myrrha.
Several plant constituents were found to have anti-bacterial activity and/or
antifungal properties (Oliver-Beaver, 1986). These include
e.g., phenol from Acardium occidentale, quinones from Drosera indica,
alkaloid from Argemone mexicana, flavinoids from Conscora decusta
and terpinoids from Borreria verticillala. The sesquiterpene (Furano
seco-A-ring sesquiterpene curzerenone and other sesquiterpene mixtures)
which were isolated from Commiphora molmol were found to be responsible
of the activity against S. aureus (Hsieh et al.,
1998; Gibbons, 2004). The minimum inhibitory concentration
was found to be 0.7 μg mL-1 and this concentration compared
very well with ciprofloxacin activity against the same strain (Dolara
et al., 2000). Tucker (1986) earlier claimed
that myrrh was used for treating wounds and as a local eye medication, which
was probably due to the activity of sesquiterpenes present in myrrh. Hsieh
et al. (1998) and Dolara et al. (2000)
investigations are in line with what has been reported in the present investigation.
However, the minimum inhibitory concentration for the ether extract in the present
study was 10 mg mL-1 whereas the ethanolic extract was 40 mg mL-1.
This finding suggests that the sesquiterpenes concentration is higher in the
ether extracts of C. myrrha compared to the ethanolic extracts. Unlike
what has been reported in the present study, Kubmarawa et
al. (2007) in Nigeria, reported activity of Commiphora kerstingii
extracts against C. albicans, B. subtilis and E. coli but
not against S. aureus and P. aeruginosa. This is probably due
to difference in species and geographical region. Ali et
al. (2008) and Suleiman et al. (2010)
evaluated the oil extracts from Commiphora kua from Socotra and Commiphora
harveyi extracts against the phyopathogenic fungus Cladosporium cucumerinum
and the animal pathogen Cryptococcus neoformans respectively, in both
studies antifungal activities were reported and this is support with what we
found in case of C. albicans. These findings are in support to our study
indicating the antifungal activity of Commiphora sp.
This study confirms that ethanolic and ether extracts of C. myrrha possess in vitro antibacterial and antifungal activity. In case such extracts are to be used for food preservation or medicinal purposes, issues of safety and toxicity must be taken into account. However, further investigation is needed in this important area to explain the active constituents present in this plant extract in inhibiting bacterial and fungal growths.
The authors are grateful to Highness Prince Bandar bin Saud, Secretary General of the Saudi Wildlife Commission, for encouraging and supporting research at King Khalid Wildlife Research Centre and to Dr Thomas Butynski, KKWRC Director, for encouragement and support.
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