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Journal of Medicinal Plants Research Vol. 5(1), pp. 7-14, 4 January, 2011
Available online at http://www.academicjournals.org/JMPR
ISSN 1996-0875 ©2010 Academic Journals
Full Length Research Paper
Bacteriostatic and bactericidal activities of
Andrographis paniculata extracts on skin disease
causing pathogenic bacteria
Abubakar Sule1*, Qamar U. Ahmed2, Othman A. Samah1 and Muhammad N. Omar1
1Department of Biomedical Sciences, Faculty of Science, International Islamic University Malaysia (IIUM), 25200
Kuantan, Pahang DM, Malaysia.
2Department of Pharmaceutical Chemistry, Faculty of Pharmacy, International Islamic University Malaysia (IIUM), 25200
Kuantan, Pahang DM, Malaysia.
Accepted 10 May, 2010
Non-polar (dichloromethane) and polar (MeOH and aqueous) extracts of Andrographis paniculata
(whole plant) were evaluated for in vitro antibacterial activity against 10 skin disease causing bacterial
strains (6 gram positive strains; Staphylococcus saprophyticus, Staphylococcus epidermis,
Staphylococcus aureus, Streptococcus pyogenes, Bacillus anthracis, Micrococcus luteus) and 4 gram
negative strains (Proteus mirabilis, Proteus vulgaris, Neisseria meningitis, Pseudomonas aeruginosa)
using disc diffusion method at three different concentrations; 1000, 500 and 250 µg/disc respectively.
The extracts showed significant antibacterial activities against both Gram-positive and Gram-negative
bacterial strains tested. Highest significant antibacterial activity was exerted by the aqueous extract
against M. luteus at 1000 µg/disc and the least activity was exhibited by the DCM extract against N.
meningitis at 250 µg/disc. The minimum inhibitory concentration (MIC) and minimum bactericidal
concentration (MBC) observed were between 150 to 300 µg/ml and 250 to 400 µg/ml respectively,
depending on microorganism and the nature of various extracts. Time-kill experiments indicated that A.
paniculata extracts have bactericidal characteristic against most of the Gram positive bacteria and
bacteriostatic activity against both Gram negative and Gram positive bacteria. These results candidly
suggest the presence of promising antibacterial substances in the polar as well as non-polar extracts
which could be the source of potential phytomedicine for the treatment of skin infections caused by the
pathogenic bacterial strains. Our findings explicitly support its traditional claims and form a strong
basis for further sincere efforts to explore A. paniculata’s antibacterial potential to treat skin frailties
efficaciously.
Key words: Andrographis paniculata, antimicrobial activity, skin infections, disc diffusion method, minimum
inhibitory concentration, minimum bactericidal concentration, time to kill assay.
INTRODUCTION
There have been high rise in the frequency of certain skin
infections in developing countries including Malaysia.
Indeed, skin infections are among the most prevalent in
the world. Bacterial skin infections are common
outpatient problems and the 28th most common
*Corresponding author. E-mail: boukar74sule@yahoo.com. Tel:
+60175935594.
infections diagnosis in hospitalized patients (Elixhauser et
al., 2001). Studies have stated that it may account for up
to 17% of clinical visits (Sadick, 1997). Therapies of
bacterial skin infections are frequent problems due to the
emergence of resistant bacterial strains to numerous
antibiotics (Marimoto et al., 1999). Some plants have
shown the ability to overcome resistance in some
organisms and this has led to researchers’ investigating
their mechanisms of action and isolating active
compounds from them (Ncube et al., 2007). Nowadays,
8 J. Med. Plant. Res.
researches on medicinal plants have attracted a lot of
attention globally. A number of evidences have been
accumulated to demonstrate the promising potentials of
medicinal plants used in various traditional,
complementary and alternative systems (Kanokwan et
al., 2008).
Andrographis paniculata (Burm.f.) Wall. ex Nees.,
belongs to the family of Acanthaceae and is popular
worldwide with the name of King of Bitters in English and
Hempedu bumi in Malay. It is an annual herbaceous plant
which is widely cultivated in southern Asia, Scandinavia,
China and some parts of Europe. The leaves and roots
have traditionally been used over the centuries in Asia
and Europe as a folklore medicine for a wide variety of
ailments or as herbal supplements for health promotion.
In traditional Chinese medicine, it is widely used to get rid
of body heat, as in fevers and to dispel toxins from the
body. In Scandinavian countries, it is commonly used to
prevent and treat common cold (Caceras et al., 1997).
Previous studies have explicitly revealed that A.
paniculata has a wide range of pharmacological effects
and some of them extremely beneficial such as anti-
inflammatory (Shen et al., 2002), anti-diabetes (Syahrin
et al., 2006), antidiarrhoeal (Gupta et al., 1990), antiviral
(Wiart et al., 2005), antimalarial (Rahman et al., 1999),
hepatoprotective (Trivedi and Rawal, 2005), anticancer
(Cheung et al., 2005; Tan et al., 2005; Zhou et al., 2006),
antihuman immunodeficiency virus (HIV) (Calabrese et
al., 2000), immune stimulatory (Iruretagoyena et al.,
2005), antimicrobial (Prajjal et al., 2003; Coon et al.,
2004; Limsong et al., 2004) and antisnakebite activity
(Samy et al., 2008). Diterpenoids and flavonoids are the
main chemical constituents of A. paniculata which are
believed to be responsible for the most biological
activities of this plant (Tang and Eisenbrand, 1992).
A. paniculata has been used in the treatment of some
skin infections in India and China by folkloric medicine
practitioners. It is considered beneficial to the skin and is
used both internally and externally for this purpose (Jain,
1991). Evidences on its wide use by the traditional clerics
in treating some infections of the skin (Tapsell et al.,
2006) have prompted us to choose and confirm this plant
for further evaluation in order to ascertain its antibacterial
potential to treat skin infection that are caused by some
pathogenic bacterial strains.
METHODOLOGY
Collection and preparation of plant material
Fresh plant material (5 kg) of A. paniculata was procured from the
botanical gardens of the Forest Research Institute of Malaysia
(FRIM), Kuala Lumpur, Malaysia. Specimen s ample was
authenticated by Dr. Saw Leng Guan (Taxonomist, FRIM) and
deposited (voucher specimen number: NMPC-KOS-025) in the
Herbarium, Kulliyyah of Pharmacy, IIUM, Malaysia. All parts of the
plant material were dried in a protech laboratory dryer (LDD-720) at
37°C in the dark for 5 days and pulverized to powdered form using
the Fritsch Universal Cutting Mill. This was then stored in a
desiccators at 2°C until further use.
Materials
All the chemicals and standard antibiotics were purchased from
Fisher scientific chemicals, UK and Oxoid Ltd England r espectively.
All the solvents used were of analytical grade. Precoated silica gel
60 F254 TLC plates were purchased from Merck, Germany.
Preparation of non-polar and polar extracts
500 g dry powder of A. paniculata (whole plant) was sequentially
extracted with dichloromethane and methanol using the Soxhlet
apparatus on the water bath for 12 h each (Harborne, 1998). Each
of the extracts was carefully filtered using filter paper (Whatman No.
A-3) and concentrated using a rotary evaporator (Buchi Rotary
Evaporator, R-210) at 40°C. The final concentrated extracts were
stored at -18°C in labeled sterile bottles and kept as aliquots until
further evaluation.
Another 500 g of powdered sample of the herb was extracted by
soaking in 1 L double distilled water in a round bottom flask, stirred
for about 6 min, closed tight using a rubber c ork and left overnight
at room temperature. Thereafter, the s olution was filtered using filter
paper (Whatman No. A-1) and extract was freeze dried and
carefully stored at -18°C in labeled sterile bottles.
Microorganisms
Ten skin disease causing bacterial strains were taken into
consideration, viz., (6 Gram-positive: Staphylococcus saprophyticus
(IMR S-1242), Staphylococcus epidermis (IMR S-947),
Staphylococcus aureus (IMR S-277), Streptococcus pyogenes (IMR
S-526), Bacillus anthracis (IMR B-132), Micrococcus luteus (IMR B-
7) and 4 gram negative strains: Proteus mirabilis (IMR P-76),
Proteus vulgaris (IMR P-147), Neisseria meningitis (IMR N-349),
Pseudomonas aeruginosa (IMR P-84)). All bacterial strains were
obtained directly from the Institute for Medical Research (IMR),
Kuala Lumpur, Malaysia. The test organisms were sub-cultured at
37°C for 24 h and maintained on nutrient agar media.
Antibacterial activity screening
The agar disc diffusion method was employed for the determination
of antibacterial activities of the polar (MeOH and aqueous) and non-
polar (DCM-dichloromethane) extracts of A. paniculata (NCCLS,
2004). 6 Gram-positive (S. saprophyticus, S. epidermis, S. aureus,
S. pyogenes, B. anthracis, M. luteus) and 4 Gram-negative (P.
mirabilis, P. vulgaris, N. meningitis, P. aeruginosa) st andard
bacterial strains of human skin disorders were used. All bacterial
cultures were first grown on nutrient agar plates at 37°C for 24 h.
Few colonies (2 to 3) of similar morphology of the respective
bacteria were transferred to a liquid medium (Mueller Hinton Broth)
and incubated until adequate growth of turbidity equivalent to
McFarland 0.5 turbidity standard was obtained. The inocula of the
respective bacteria were streaked on to the Mueller Hinton plates.
The dried plant extracts were dissolved in 10% aqueous dimethyl
sulfoxide (DMSO) and sterilized by filtration through a 0.45 mm
membrane filter. Sterile f ilter paper discs (5 mm) (W hatman no. 1)
were punched and impregnated with 10 µl of the DCM, MeOH and
aqueous extracts (c orresponding to 1000, 500 and 250 µg/disc) and
allowed to dry at room temperature. These were placed on the
Mueller-Hinton Agar plates inoculated with the t est strains. The
plates were then allowed to stay for 1 h at room temperature and
finally incubated at 37°C for 24 h (Heraeus GmbH, D-6450, and
Germany). The assessment of antibacterial activity was based on
the measurement of diameter of inhibition zone (mm) formed
around the disc. Antibacterial activity was assigned by measuring
the inhibition zone formed around the discs. The experiment was
done three times and the mean values were presented.
Tetracycline (30 µ g) and Gentamicin (30 µg) were used as positive
controls while 10% DMSO was taken as negative control.
Determination of minimum inhibitory concentration (MIC)
The MIC of the crude extracts of A. paniculata was determined by
agar dilution method as validated by EUCAST, 2000. The growth
media, Mueller Hinton Agar were first prepared and sterilized by
autoclaving (W ebco GmbH and Co. KG Bad Schwartau, Germany).
The sterilized media was allowed to cool to 50°C and 18 ml of the
molten agar was added to test tubes which contained 2 ml of
different concentration of the test crude extract and the control
(10% DMSO). The mixture of the media and the test crude extracts
were thoroughly mixed and poured into pre-labeled sterile petri-
dishes on a level surface. Additional petri-dishes containing only the
growth media were prepared in the same way so as to serve for
comparison of growth of the respective organisms. The
concentrations of the extracts used in this test ranged from 150 to
300 µg/ml. The plates were then set at room temperature and dried.
The suspensions of the respective microorganisms having density
adjusted t o 0.5 McFarland turbidity standards were inoculated onto
the series of agar plates. Three loopful of the suspension were
transferred into each plate and spread evenly. The plates were then
incubated at 37°C for 24 h. The lowest concentration of the extract
which inhibited the growth of the respective organisms was taken
as MIC.
Determination of minimum bactericidal concentration (MBC)
The MBC was determined by the micro broth dilution method
(NCCLS, 2003, 2004). Plant extracts were resuspended in 10%
DMSO (which had no activity against t est microorganisms) to make
final concentration of 500 µg/ml, this was then serially diluted by
adding to the broth media in a 96-wells microtiter plates to obtain
450, 400, 350, 300, 250, 200,150 and100 µg/ml. Thereafter, 100 µl
inoculum was added into each well. Bacterial suspensions were
used as negative control, while broth containing standard drugs
(Tetracycline and Gentamicin) were used as positive control. The
micro titer plates were incubated at 37°C for 24 h. Each extract was
assayed in triplicates and each time two s ets of micro plates were
prepared, one was kept for incubation while another set was kept at
4°C for comparing the turbidity in the wells of micro plate. The
turbidity of the wells in the micro titer plate was interpreted as
visible growth of microorganisms. The MBC was determined by sub
culturing 50 µl from each well showing no apparent growth. Least
concentration of extract showing no visible growth on subculturing
was taken as MBC.
Time to kill assay
Time-kill assay were performed as described by Ernst et al. (2002)
with little modifications to determine the rate of killing of the chosen
bacteria by t he extracts of A. paniculata. 1000 µg/ml concentrations
of each extract were prepared. Inoculums of the test microganisms
were prepared from 24 h cultures grown on Mueller-Hinton Broth
(MHB) and suspensions adjust ed to a turbidity equivalent to
108CFU/ml. Equal volumes of the diluted inoculums and the
extracts to be tested were mixed and incubated at 37°C. At time
intervals of 2 to 24 h, 50 µl of the mixed suspension was spread on
two separate nutrient agar plates and incubated for 24 h at 37°C.
The mean number of colonies were obtained and compared with
that of control in which the plant extracts was replaced with DMSO.
Sule et al. 9
Each experiment was repeated thrice. The colony count results
were expressed as a percentage of control (that is, percentage of
bacteria killed compared to the control).
Phytochemical screening
Phytochemical screening of plant extracts was carried out
qualitatively for the presence of alkaloids, terpenoids, tannins,
flavonoids, saponins, cardiac glycosides and steroids. Alkaloid
detection was carried out by extracting 1 g powdered sample with 5
ml methanol and 5 ml of 2N HCl; and then treating the filtrate with
Meyer's and Wagner's reagents. The samples were scored positive
on the basis of turbidity or precipitation. Flavonoids were t ested by
heating 1 g powdered sample with 10 ml ethyl acetate over a steam
bath (40 to 50°C) for 5 min; filtrate was tr eated with 1 ml dilute
ammonia. A yellow c olouration demonstrated positive test for
flavonoids. The presence of tannins was confirmed by boiling 0.5 g
powdered sample in 20 ml distilled water, followed by addition of 3
drops of 5% FeCl3 to the filtrate. Development of brownish-green or
blue black colouration was t aken as positive f or the presence of
tannins. Saponins content was determined by boiling 1 g powdered
sample in 10 ml distilled water for 15 min and after cooling, the
extract was shaken vigorously to record froth formation. Cardiac
glycosides were identified by extracting 2 g sample in 10 ml
methanol. Five ml of this methanolic extract was treated with 2 ml
glacial acetic acid containing 1 drop of 5% FeCl3 solution. This
solution was carefully transferred to surface of 1 ml conc. H2SO4.
The formation of reddish brown ring at the junction of two liquids
was indicative of cardenolides/cardiac glycosides (Harborne, 1998).
Statistical analysis
All tests were conducted in triplicates. All values have been
expressed as mean ± standard deviation and the c omparison of the
antibacterial activity of the samples with standard antibiotics was
evaluated by applying t-test. P ≤ 0.05 values were considered to
indicate statistically significant difference.
RESULTS AND DISCUSSION
The results of present study are encouraging as all the
tested extracts revealed antibacterial potential, although
the inhibitory activity was strain specific and
concentration dependant. The dichloromethane,
methanolic and aqueous extracts of A. paniculata were
assessed at 3 different concentrations by using disc
diffusion method against 10 bacterial strains notable for
causing chronic skin infections and expressed as the
average diameter of the zone of inhibition of bacterial
growth around the disc. The MIC and the MBC of active
extracts were determined by the agar dilution and micro
broth dilution assays respectively. The extracts displayed
relative antibacterial activity against most of the tested
microorganisms with the diameter of inhibition zones
ranging between 6.00 ± 1.00 to 23.17 ± 0.76mm (Table
1). The Gram-positive strains used for this study were the
most susceptible to growth inhibition by the plant extracts
forming zones of inhibition ranging from 7.00 ± 0.00 to
23.17± 0.76mm. The DCM extract was found to exhibit
the least potent antibacterial activity against
10 J. Med. Plant. Res.
Table 1. Antibacterial activity of polar and non-polar extracts of A. paniculata. Numbers indicate the mean diameters of inhibition of triplicate experiments ± (SD).
Zone of inhibition diameters in mm
Extracts DCM MeOH Aqueous Gentamicin Tetracycline
Concentration 1000 µg/disc 500 µg/disc 250 µg/disc 1000 µg/disc 500 µg/disc 250 µg/disc 1000 µg/disc 500 µg/disc 250 µg/disc 30 µg 30 µg
S.saprophyticus IMR S-1242 19.33 ± 1.15 16.50 ± 0.87 7.00 ± 0.00 22.00 ± 1.53 18.83 ± 0.76 0.00 ± 0.00 20.67 ± 1.15 16.33 ± 1.53 8.33 ± 0.76 24.33 ± 1.52 0.00 ± 0.00
S. epidermis IMR S-947 18.00 ± 0.50 18.00 ± 0.50 8.33 ± 1.04 20.67 ± 0.58 18.67 ± 1.15 8.00 ± 1.00 19.00 ± 0.00 17.67 ± 0.58 12.00 ± 0.50 18.67 ± 1.52 0.00 ± 0.00
S. aureus IMR S-277 20.00 ± 1.50 17.00 ± 0.00 14.00 ± 0.50 22.00 ± 0.00 17.50 ± 0.50 13.50 ± 0.87 19.00 ± 0.00 15.67 ± 0.58 10.00 ± 1.00 22.33 ± 1.08 0.00 ± 0.00
B. anthracis IMR B-132 17.83 ± 0.76 13.33 ± 2.08 0.00 ± 0.00 20.00 ± 1.00 17.33 ± 0.58 14.83 ± 0.76 16.67 ± 1.15 10.00 ± 1.00 8.33 ± 0.58 21.67 ± 1.51 14.33 ± 1.21
M. luteus IMR B-7 17.67 ± 1.73 14.33 ± 1.53 0.00 ± 0.00 19.33 ± 0.58 15.50 ± 1.00 13.33 ± 0.76 23.17 ± 0.76 21.00 ± 0.00 0.00 ± 0.00 14.00 ± 0.00 21.67 ± 0.52
S. pyogenes IMR S-526 17.67 ± 1.15 11.17 ± 0.76 0.00 ± 0.00 17.00 ± 0.00 13.33 ± 0.00 9.00 ± 0.00 22.67 ± 0.58 16.33 ± 0.00 0.00 ± 0.00 17.67 ± 1.73 21.33 ± 1.06
P. mirabilis IMR IMR P-76 18.33 ± 0.76 14.00 ± 1.00 6.67 ± 0.58 20.00 ± 0.00 17.00 ± 1.00 16.67 ± 0.58 19.00 ± 0.00 18.33 ± 0.78 15.33 ± 1.53 29.33 ± 0.89 27.00 ± 0.00
P. vulgaris IMR P-147 16.33 ± 0.58 15.33 ± 1.53 12.00 ± 0.00 19.00 ± 0.50 16.33 ± 1.04 7.00 ± 0.00 18.67 ± 1.53 16.33 ± 0.58 15.00 ± 0.00 19.33 ± 1.30 18.67 ± 1.05
P. aeruginosa IMR P-84 13.33 ± 0.58 13.33 ± 0.58 0.00 ± 0.00 10.33 ± 1.04 9.00 ± 1.73 7.00 ± 0.00 15.00 ± 0.00 11.00 ± 0.00 11.17 ± 1.04 20.33 ± 1.52 0.00 ± 0.00
N. meningitis IMR N-349 17.67 ± 0.76 15.33 ± 0.58 10.00 ± 0.00 18.33 ± 0.58 9.67 ± 0.58 7.00 ± 1.00 12.17 ± 0.76 14.67 ± 0.58 6.00 ± 0.00 12.00 ± 0.00 18.67 ± 0.52
S. saprophyticus (7.00 ± 0.00mm) at 250 µg/disc
and the aqueous extract displayed the most
potent activity against M. luteus (23.17 ± 0.76
mm) at 1000 µg/disc (Figure 1). The highest MIC
value was observed at 300 µg /ml exerted by the
aqueous extract against M. luteus, DCM extract
against M. lutues and B. anthracis respectively.
The least MIC was 150 µg/ml exerted by the
aqueous extract against S. aureus and the
methanolic extract on B. anthracis
respectively.The highest MBC was observed at
400 µg/ml exerted by the DCM extract against S.
pyogenes and aqueous extract against S.
saprophyticus and S. epidermis.The least MBC
was 250 µg/ml exerted by the DCM extract
against S. saprophyticus and the methanolic
extract against M. luteus and S. pyogenes (Table
3). However, no activity was observed with the
DCM, methanolic and aqueous extracts of the
plant at 250 µg/disc against M. luteus, S.
pyogenes and S. saprophyticus. The Gram
negative strains were less sensitive to the plant
extracts as compared to the Gram positive,
forming zones of inhibition ranging from 6.00 ±
1.00 to 20.00 ± 1.26mm. The aqueous extract was
found to be the least potent against N. meningitis
(6.00 ± 1.00mm) at 250 µg/disc and the
methanolic extract showed the most potent
activity against P. mirabilis (20.00 ± 1.26mm) at
1000 µg/disc (Figure 2). The highest MIC value
was found to be 300 µg/ml exerted by the DCM
extract against P. aeroginosa and the least was
150 µg/ml exerted by the aqueous extract against
P. vulgaris and the methanolic extract extract on
N. meningitis respectively (Table 2). MBC values
were undetected against all gram negative strains
tested (Table 3). No activity was observed with
the DCM, methanolic and aqueous extracts of the
plant at 250 µg /disc against P. aeroginosa. P
aeroginosa and all Staphylococcus strains used
for the study were found to be resistant to
tetracycline (Table 1).
All tested extracts of A. paniculata exhibited
bacteriostatic action against both Gram positive
and negative bacterial strains (MIC) but revealed
bactericidal action against Gram positive strains
only except for B. anthracis (MBC). The colony
count method was used to ascertain the time
taken for the extracts to completely kill the
bacterial cell. The strains which displayed
considerably good sensitivity to plant extracts
were selected further to determine bactericidal/
bacteriostatic activities by the time-kill assay viz.,
M. luteus (Gram positive) and P. mirabilis (Gram
Negative); the results explicitly indicated a
significant decrease in mean colony count of the
tested strains as compared to the control at each
time interval leading to the complete killing of M.
luteus cells by the aqueous extract at 12 h, the
MeOH extract at 14 h and the DCM extract at 20 h
respectively, but these extracts were unable to kill
the cells of P. mirabilis totally even though there
was much decrease in mean colony count, P.
mirabilis cells were still viable after 24 h. The
results indicated that all extracts were bactericidal
against M. luteus but exhibited bacteriostatic
action against P. mirabilis (Figures 3 and 4).
Phytochemical screening of plant extracts was
carried out qualitatively for the presence of
terpenoids, tannins, flavonoids, saponins, cardiac
glycosides and steroids (Harborne, 1998). All
extracts showed the presence of terpenoidal and
Sule et al. 11
Gram positive bacteria
Zones of inhibition (mm)
Figure 1. Zones of inhibition (mm) of the DCM, MeOH and aqueous extracts of A. paniculata at 1000 µg/disc
on Gram positive bacterial strains.
Gram
negative
bacteria
Zones of inhibition (mm)
Figure 2. Zones of Inhibition (mm) of the DCM, MeOH and aqueous extracts of A. paniculata at 1000
µg/disc on Gram negative bacterial str ains.
flavonoidal compounds. However, MeOH extract gave
positive test for all compounds taken into consideration
and showed the maximum number of aforementioned
compounds on precoated silica gel 60 F254 TLC plate
which could be responsible for its strong antibacterial
activity against all microorganisms taken into account.
In the present era, plant and herb resources are
abundant, but these resources are dwindling fast due to
the onward march of civilization (Vogel, 1991). Although,
a significant number of studies have been used to obtain
purified phytochemicals, very few screening programmes
have been initiated on crude plant materials. It has also
12 J. Med. Plant. Res.
Table 2. MIC of the plant extracts on the bacterial strains.
Minimum inhibitory concentrations (MIC) (µg/ml)
Bacterial strains Dichloromethane Methanol Aqueous
Gram positive strains
S. saprophyticus 150 250 200
S. epidermis 200 250 250
S. aureus 250 200 150
B. anthracis 300 150 200
M. luteus 300 250 300
S. pyogenes 200 250 250
Gram negative strains
P. mirabilis 200 250 250
P. vulgaris 250 250 150
P. aeruginosa 300 250 250
N. meningitis 200 150 250
Table 3. MBC of the plant extracts on the bacterial strains.
Minimum bactericidal concentrations (MBC) (µg/ml)
Bacterial strains Dichloromethane Methanol Aqueous
Gram positive
S. saprophyticus 250 300 400
S. epidermis 350 300 400
S. aureus 400 300 300
B. anthracis UD UD UD
M. luteus 350 250 350
S. pyogenes 400 250 350
Gram negative
P. mirabilis UD* UD UD
P. vulgaris UD UD UD
P. aeruginosa UD UD UD
N. meningitis UD UD UD
*UD=Undetected.
been widely observed and accepted that the medicinal
value of plants lies in the bioactive phytocomponents
present in the plants (Veeramuthu et al., 2006). The
greater susceptibility of Gram-positive bacteria to plant
extracts has been previously reported in South American
(Paz et al., 1995), African (Kudi et al., 1999; Vlietinck et
al., 1995) and Australian (Palombo and Semple, 2001)
medicinal plant extracts. Susceptibility differences
between Gram-positive and Gram-negative bacteria may
be due to cell wall structural differences between these
classes of bacteria. The Gram-negative bacterial cell wall
outer membrane appears to act as a barrier to many
substances including antibiotics (Tortora et al., 2001;
Gurinder and Daljit, 2009). The significant results
obtained in our study confirm the antibacterial potential of
the plant investigated, and its usefulness in the treatment
of skin infections. This in vitro study corroborates the
antibacterial activity of A. paniculata used in folkloric
medicine to treat skin infections (Jain, 1991; Ahmed et
al., 1998). All these extracts were manifested to exhibit
inhibitory activity against most of the pathogenic bacteria
which cause chronic bacterial skin infections. However,
they were ineffective at low concentrations against S.
saprophyticus, B. anthracis, M. luteus, S. pyogenes, and
P. aeroginosa. Hence, their medicinal uses in infections
associated with these bacterial species are not
recommended. Antibacterial efficacy shown by these
extracts provides a scientific basis and thus, validates
their traditional uses as homemade remedies in the
treatment of skin infirmities which are associated with
these bacteria. The present study revealed that the crude
polar and non-polar extracts contain a number of
phytoconstituents whose isolation and purification may
yield significant novel antimicrobial agents. Further,
Sule et al. 13
Time (h)
Colony count (percentage of control
)
Aqueous
Figure 3. Colony count of M. luteus with A. paniclata MeOH, DCM and aqueous
extracts.
Time (h)
Colony count (percentage of control
Aqueous
Figure 4. Colony count of P. mirabilis with A. paniclata MeOH, DCM and aqueous
extracts.
investigation to obtain information on chemical composi-
tion, to purify and determine the structures of active
principles in A. paniculata have been in progress in our
laboratory.
ACKNOWLEDGEMENT
The authors are extremely grateful to the Faculty of
Science and Faculty of Pharmacy, International Islamic
14 J. Med. Plant. Res.
University Malaysia (IIUM), 25200 Kuantan, Pahang DM,
Malaysia, for providing all research facilities to
accomplish this study. Also, a big thanks to Research
Management Center, IIUM for furnishing grant (EDW B
0904-267) to carry out this work effectively.
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