Antimicrobial Evaluation of Buddleja asiatica Lour. Leaves and Flowers Extract
to cite this article:
Shivani Joshi, Devendra Mishra, K.S. Khetwal and Ganga Bisht, 2012. Antimicrobial Evaluation of Buddleja asiatica Lour. Leaves and Flowers Extract. Research Journal of Phytochemistry, 6: 92-95.
Received: November 02, 2011;
Accepted: February 20, 2012;
Published: March 26, 2012
Medicinal plants contain various biologically active compounds which are known
to possess antimicrobial properties (Cowan, 1999). These
plant based antimicrobials have been documented as good source of medicines
with enormous therapeutic potential. Many reports have confirmed the potential
of these antimicrobials in the prevention of some infectious diseases with no
side effects, as are often related with synthetic antimicrobials (Iwu
et al., 1999). They are also responsible for the medicinal plants
to be used as food preservatives (Sunilson et al.,
2009). Developing countries still depend mainly on medicinal herbs due to
their cheaper cost and their effectiveness in the treatment of various infectious
diseases with lesser side effects (Butkhup and Samappito,
2011; Karim et al., 2011). Traditional tropical
herbs could serve as good source of new safe, biodegradable and renewable drugs
for the treatment of fungal or related ailments (Musyimi
and Ogur, 2008).
In recent times the critical area of primary health concern is the usual causative
agents that are responsible for the incidence of new and re-emerging infectious
diseases. They have now been increasingly resistant to some or most existing
antibiotics due to development of resistant strains (Khanahmadi
et al., 2010; Bonjar, 2004). Therefore, the
search for new antimicrobial compounds with novel action mechanisms becomes
B. asiatica (Buddlejaceae), commonly known as butterfly bush has been
used as an abortifacient and in the treatment of skin complaints (Chopra
et al., 1986; Anonymous, 1993). The present
study was carried out to investigate the antimicrobial potential of crude methanolic
extract of leaves and flowers of B. asiatica Lour. occurring in Kumaun
Himalayas, India, against a wide range of pathogenic bacterial and fungal strains
which have not been evaluated in the previous studies.
MATERIALS AND METHODS
Plant material: The leaves and flowers of the plant were collected in the month of February, 2009 from Nainital, India and authenticated by Botanical Survey of India, Dehradun. A voucher specimen (No. 112965) was deposited in the Herbarium Section at BSI, Dehradun, India.
The leaves and flowers of the plant were shade dried, powdered and extracted with methanol using Soxhlet apparatus. After extraction the filtrate was concentrated on a rotary evaporator under vacuum at 20°C till a residual mass was obtained.
Phytochemical screening: Phytochemical screening of methanolic extract
of leaves and flowers of B. asiatica, showed the presence of triterpenoids,
steroids, flavonoids and saponins (Harborne, 1984).
Microorganisms: Three gram positive, three gram negative and two fungi were used for antimicrobial activity studies. Gram positive bacteria were S. aureus (MTCC 3160), B. subtilis (MTCC 441) and B. mycoides (MTCC 645). Gram negative bacteria were E. coli (MTCC 406), P. aeruginosa (MTCC 424) and P. vulgaris (MTCC 426). Yeast like fungi used were C. albicans (MTCC 227) and A. niger (MTCC 404). Required microorganisms were procured from Institute of Microbial Technology, Chandigarh, India.
Determination of zone of inhibition: Antimicrobial tests of selected
microoganisms were carried out by disc diffusion method (Murray
et al., 1995). Nutrient agar plates and potato dextrose agar plates
were prepared for antibacterial and antifungal activity respectively. The test
solutions of the methanol extract (4000, 2000, 1000, 500, 250 μg mL-1)
were prepared by dissolving the extract in dimethylsulphoxide (DMSO). The plates
were cooled down at 20°C and then inoculated with bacterial and fungal cultures
by spreading the inoculum over the entire agar surface. The filter paper discs
(5 mm in diameter, Whatman filter paper 1) were individually impregnated with
0.1 mL of the test solutions which were subsequently placed on the surface of
the inoculated petri dishes. Chloramphenicol (25 μg disc-1),
ampicillin (25 μg disc-1) and fluconazole (25 μg mL-1)
were used as positive controls. As a negative control, a blank disc impregnated
with DMSO (20%) was used. The test discs, standard discs and blank discs were
placed in petridish with a particular microorganism. The petridishes were then
incubated at 37°C for 24 h for bacterial growth and at 27°C for 48 h
for the growth of yeast. After 24 h of incubation, the diameter was observed
for zone of inhibition (measured in mm including the disc size). Each extract
was analyzed in triplicate and observed values of ZOI were then expressed as
RESULTS AND DISCUSSION
Antimicrobial potential of the extracts were evaluated by measuring the diameter
of zones of inhibition (mm), including the diameter of disc and the results
(mm of zone of inhibition) were expressed as average values. The zone of inhibition
markedly decreased on decreasing the concentration of the extracts for all the
strains used for study. The results of B. asiatica antibacterial evaluation
showed that B. subtilis, B. mycoides and P. vulgaris were insensitive
for both leaves and flowers extract. The best antibacterial activity was shown
by flowers extract against P. aeruginosa while leaves extract showed
good activity against S. aureus.
|| Antimicrobial activity of methanol extract of Buddleja
asiatica L. leaves and flowers
|AM: Ampicillin (25 μg disc-1), CP: Chloramphenicol
(25 μg disc-1), FZ: Fluconazole (25 μg mL-1)
In case of antifungal activity, leaves extract was found inactive against
A. niger while flowers extract showed very good activity. C. albicans
was found sensitive for both the extracts. Antimicrobial activity of methanolic
extract of leaves and flowers of B. asiatica could be due to the presence
of triterpenoids, steroids, polyphenols, flavonoids (Taleb-Contini
et al., 2003; Jain et al., 2001;
Okoro et al., 2010). The antibacterial activity
of flavonoids and polyphenolic compounds might be due to their ability to complex
with bacterial cell wall and therefore, inhibiting the microbial growth (Sivapriya
et al., 2011).
The result of screened plant extracts for antibacterial and antifungal activity
is summarized in Table 1. The results clearly indicate that
the antibacterial and antifungal activity vary with the type of tested microbes.
This is based upon the fact that antimicrobial activity of plant extracts depends
on the species of the plant and the type of tested microorganisms (Obeidat
et al., 2012). The present study could be used for further investigation
on this plant to find the role in the total remedies of a wide range of microbial
diseases of plants and animals. The studied plant flowers may be a good source
of future drugs that could be used in the treatment of infections caused by
From the results, it can be concluded that both the extracts were able to inhibit the growth of the investigated bacterial strains; the most susceptible bacteria was P. aeruginosa while resistant were B. subtilis, B. mycoides and P. vulgaris. Since B. asiatica flowers demonstrated good activity against the antibiotic low susceptibility pathogen, P. aeruginosa, the plant can be used in the treatment of infections caused by these microbes.
The authors wish to thank Botanical Survey of India, Dehradun for plant identification and Department of Pharmaceutical Sciences, Kumaun University, for providing necessary facilities to carry out present investigation.
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