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Journal of Ethnopharmacology 116 (2008) 102–111

Antibacterial activity of plants used in traditional medicines

of Ghana with particular reference to MRSA
George A. Pesewu a,b , Ronald R. Cutler a,∗ , David P. Humber a
aSchool of Health and Bioscience, University of East London, Stratford, London E15 4LZ, UK
b Centre for Scientific Research into Plant Medicine (CSRPM), P.O. Box 73, Mampong-Akwapim, Ghana
Received 20 June 2006; received in revised form 7 November 2007; accepted 8 November 2007
Available online 17 November 2007

Abstract
Ethnopharmacological relevance: : In an ethno botanical survey carried out in the Akwapim-North district of the Republic of Ghana, 25 plant
species, used in traditional medicine to treat skin disease and/or to treat antimicrobial (viral, bacterial or protozoan) infections were identified.
Aim of Study: : To investigate the antimicrobial activity of traditional Ghanaian medicines with special interest in anti-methicillin-resistant
Staphylococcus aureus (MRSA) activity.
Materials and methods: : Chloroform, ethanol and aqueous extracts (including use of a Stomacher) of these plants were prepared and agar-well
diffusion tests, MIC’s and MBC’s were used to investigate antimicrobial activity.
Results: Extracts of 13 plant species inhibited the growth of one or more of the following bacteria: MRSA, methicillin-sensitive Staphylococcus
aureus (MSSA), Streptococcus pyogenes, Escherichia coli, Pseudomonas aeruginosa and Proteus vulgaris. Extracts from 11 of these 13 plant
species also inhibited the growth of three or more of 14 additional clinical isolates of MRSA. Aqueous extracts of Alchornea cordifolia were
active against all 21 bacterial strains tested and showed the highest levels of antibacterial activity overall with MIC’s against MRSA in the range
of 1.6–3.1 mg ml−1 and MBC’s in the range of 6.3–12.5 mg ml−1 .
Conclusions: : The presence of antibacterial activity in extracts of Elaeophorbia drupifera, Rauwolfia vomitoria and the leaves of Solanum
verbascifolium, plants traditionally used to treat skin infections, are reported for the first time. Extracts from Alchornea cordifolia, also used to
treat wounds, had the widest spectrum of antibacterial activity.
© 2007 Elsevier Ireland Ltd. All rights reserved.

Keywords: Antibacterial; MRSA; Traditional medicines; Ghana

1. Introduction
Many of the drugs currently used to treat bacterial and other
infections were first isolated from natural sources including eth-
nomedicinal plants (Coe and Anderson, 1996). Such plants may
provide new sources of therapeutic agents against multiply drug
resistant bacteria such as methicillin-resistant Staphylococcus
aureus (MRSA). MRSA is a major cause of nosocomial infec-
tion in UK hospitals and throughout the world. MRSA infections
account for one fifth of all hospital-acquired infections, costing
the UK National Health Service approximately £1 billion per
year (Cepeda et al., 2005). The problem has been aggravated by
the rapid spread and high incidence of MRSA in intensive-care
∗ Corresponding author. Tel.: +44 208 223 4162.
E-mail address: r.cutler@uel.ac.uk (R.R. Cutler).
units (Cepeda et al., 2005). The continuing rise in MRSA infec-
tion rates and its spread worldwide has led to calls for action to
control infection and develop novel anti-MRSA agents (Cutler
and Wilson, 2004; Hancock, 2007) and vaccines.
Several ethnomedicinal plant species of Ghana have
been identified and their usage documented; (Abbiw, 1990;
Anonymous, 1992; Dokosi, 1998; Mshana et al., 2000; Agbovie
et al., 2002; Anonymous, 2004). They have been used as antibac-
terial, antifungal, antiviral and antiprotozoan agents and for the
general treatment of skin diseases, dermatitis, burns, diarrhoea,
fever (pyrexia) of unknown origin, wounds, cuts, sores, coughs
and localized skin swellings.
In the present investigation, plants used in folk medicine
in the Akwapim-North district of Ghana to treat bacterial and
other skin conditions were identified and extracts were tested for
antibacterial activity. Particular attention was given to activity
against MRSA.

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 G.A. Pesewu et al. / Journal of Ethnopharmacology 116 (2008) 102–111
2. Materials and methods
2.1. Ethnobotanical survey and collection of specimens
A survey was carried out in the Akwapim-North district of
Ghana, a dry equatorial region where most of the natural vegeta-
tion has been cleared for the cultivation of food crops. It covers
an area of 544 km2 and has a population of about 105,000 that
includes the Cherepong, Larteh, and Akwamus ethnic groups.
Interviews were conducted between July and September 2002
and between November and December 2003 with 81 herbal-
ists and traditional healers in towns and villages. These villages
included Adwaso, Dawu, Kunutiase, Kwamoso, Mamfe, New
Mangoase, Obosomase, Okrakwadwo, Old Asuoyaa and Tutu.
There were three objectives to the survey; to prepare an inventory
of plants used to treat skin infections; to establish the origins,
history and overall usage of these plants and to clarify the local
procedures used to prepare and administer these medicines.
Interviews were conducted according to the procedures recom-
mended by Otshundi et al. (2000). A degree of consensus in
the listing of medicinal plants attributable to a cultural group is
important (Agelet and Valles, 2003) therefore only species with
fidelity values (percentage of interviewees citing the same plant)
of two or more were selected for this study.
Specimens of medicinal plants were collected from natural
populations. Voucher herbarium specimens were allocated to the
Ghana Herbarium (Department of Botany, University of Ghana,
Accra), the Ethnobotanical Herbarium, Centre for Scientific
Research into Plant Medicine (CSRPM), Mampong-Akwapim,
Ghana) and the School of Health and Bioscience (University of
East London, UK). Samples for laboratory investigation were
air-dried in the shade at room temperature (25–28 ◦ C), for at least
2 weeks. They were then dried at 45 ◦ C in a convection oven for
2 h to completely remove residual moisture, before milling into
fine powder. These procedures correspond to those used by the
herbalists consulted. The powders were sealed in air-tight bags
to prepare them for storage and transport.
2.2. Plant extracts
Crude extracts of the medicinal plants were obtained by both
successive extraction methods and by the use a ‘Stomacher’
Lab-Blender (A.I. Seward, UK). Extracts were prepared from
the plant organs specified by the herbalists (Table 1). Succes-
sive extractions were carried out using the procedures used at
the CSRPM when screening herbal medicines from local practi-
tioners. For each extraction, three different solvents (chloroform,
ethanol then water) were used in succession. Twenty-five grams
of dry milled plant powder was placed in a beaker (250 ml),
100 ml chloroform was added and the mixture left to soak
overnight. This mixture was then vigorously stirred for 10 min
and allowed to settle for 5 min. The supernatant liquid was
passed through filter paper (Whatman No. 1) to remove solid
plant material and the solvent evaporated from the filtrate in
vacuo at 34 ◦ C in a rotary evaporator. Finally, the residue was
dried to a constant weight in a hot air oven. This residual plant
material was further extracted, first with 80% ethanol (100 ml)
103
and then with distilled water (100 ml) using the same proce-
dure as was used for chloroform. The dry weight of each extract
was expressed as a percentage of the dry weight of the original
powdered tissue. Other workers have used successive extraction
methods with good success for obtaining antimicrobial extracts
(Camporese et al., 2003).
In parallel to the above extraction methods, aqueous extracts
were also processed using a novel stomacher (A.J Seward and
Co, UK) based method. Twenty-five grams of powdered tissue
was placed in a stomacher plastic bag, 100 ml of sterile distilled
water was added and the mixture placed in the stomacher sample
chamber. After processing for 5 min the solutions were passed
through filter paper (Whatman No. 1) and the filtrate freeze dried.
2.3. Sources of bacterial cultures
Plant extracts were tested against Escherichia coli (ATCC
25922), MSSA (methicillin susceptible Staphylococcus aureus
– NCTC 6571), MRSA (UELSHB – University of East London
School of Health and Bioscience, 102 and 103), Proteus vulgaris
(UELSHB 241), Pseudomonas aeruginosa (ATCC 27853) and
Streptococcus pyogenes (UELSHB 333). UELSHB strains are
kept at the School of Health and Bioscience, University of East
London. Tests were also carried out on 14 clinical isolates of
MRSA obtained from the Royal London Hospital courtesy of
Dr. P. Wilson.
2.4. Tests of antibacterial activity by the agar-well diffusion
method
The antibacterial activities of powdered plant extracts
were compared with controls. Tests were based on BSAC
(British Society for Antimicrobial Chemotherapy) procedures
(Andrews, 2005) modified by Cutler and Wilson (Cutler and
Wilson, 2004). Wells, 6 mm in diameter were punched in the agar
medium and powdered plant extracts dissolved in sterile distilled
water (100 (l of 50 mg ml−1 solutions) were added to each well.
As with the test solutions, 100 (l of a purified aqueous extract of
allicin at 0.5 mg ml−1 (Cutler and Wilson, 2004; Allicin Inter-
national, Rye, UK) was used in wells as an herbal antimicrobial
control. Standard paper discs (Oxoid Ltd., Basingstoke, UK),
containing gentamicin (10 (g) or vancomycin (30 (g) were used
as antibiotic controls. Plates were incubated for 24 h at 37 ◦ C
and the diameter of the inhibition zone around the test wells or
around the control discs were measured to assess antibacterial
activity. All tests were replicated three times. A pooled estimate
of the diameters of the zones of inhibition was calculated based
on the sum of squares of the deviations of all replicates from
their respective means.
2.5. Quantitative antimicrobial evaluation
The minimum inhibitory concentrations (MIC) and mini-
mum bactericidal concentrations (MBC) for the active plant
extracts were determined using standard National Committee
for Clinical Laboratory Standards methods (NCCLS, 1998).
Concentrations of plant extracts were tested in the range
 104
Table 1
Medicinal plants of the Akwapim North district of Ghana identified in the survey and used in bacteriological testsa

Family Species Voucher number Diseases treated Plant part Preparation Administration Fcb

Apocynaceae Alstonia boonei De Wild. PA2002/5 Cleansing of suppurating Stem bark Boil Oral/topical 2
wounds, open fractures
Rauwolfia vomitoria Afzel. PA2003/8 Parasitic skin diseases, Root Latex/decoction Topical 4
yaws
Asclepiadaceae Cryptolepis sanguinolenta (Lindl.) PA2002/9 Malaria Root Boil Oral 6
Schltr.

G.A. Pesewu et al. / Journal of Ethnopharmacology 116 (2008) 102–111

Parquetina nigrescens (Afzel.) Bullock PA2002/8 Candidiasis Root Poultice Topical insertion 2
Balanitaceae Balanites aegyptica (L.). Del.? PA2002/3 Herpes zoster Fruit Alcoholic extract Topical 3
Boraginaceae Heliotropium indicum L. PA2003/1 Erysipelas, thrush, Leaf Juice Topical 2
Herpes zoster
Caesalpiniaceae Cassia alata L. PA2002/1 Herpes zoster, eczema, Leaf Juice/infusion Topical 14
mycosis
Cassia podocarpa Guill. &. Perr. PA2002/15 Wounds, sore dressing, Leaf Infusion Topical 2
skin ulcers
Cassia occidentalis (Linn.) Link. PA2003/2 Guinea worm, wounds Leaf Infusion Topical 2
Capparaceae Crateva religiosa G. Forst PA2003/7 Leprosy, swollen parts of Stem bark Boil Topical 2
the body
Chenopodiaceae Chenopodium ambrasioides L. PA2002/4 Wound, skin infections Leaf Infusion Topical 5
Combretaceae Combretum micranthium G.Don. PA2002/14 Wounds, Guinea worm Root Boil Topical 3
Terminalia avicennoides Guill.&.Perr. PA2003/10 Boils Stem bark Boil Topical 2
Asteraceae Vernonia amygdalina Delile PA2002/13 Dermatitis Leaf Boil Oral/topical 5
Cucurbitaceae Momordica charantia L. PA2003/4 Measles Shoots Boil Oral/topical 2
Euphorbiaceae Alchornea cordifolia (Schum. & Thonn.) PA2002/2 Dermatitis, Herpes zoster, Leaf Juice Topical 4
Mull. Arg. ringworm, wounds
Elaeophorbia drupifera Thonn. PA2002/10 Skin infections, Guinea Leaf Boil Topical 5
worm
Ricinus communis Linn. PA2002/12 Dermatitis, keratoderma Seed Oil Topical 2
Labiatae Ocimum viride Willd. PA2002/7 Trichomoniasis Leaf Juice/poultice Topical 6
Hoslundia opposita Vahl. PA2002/11 Dermatitis Leaf Boil Oral/topical 2
Lauraceae Persea americana Mill. PA2003/6 Skin ulcers Leaf Infusion Topical 5
Myrtaceae Psidium guajava L. PA2003/9 Measles, Herpes zoster Leaf Infusion Topical 3
Rubiaceae Gardenia ternifolia Schumach. & Thonn. PA2003/3 Ulcers, syphilis, body Leaf Infusion Topical 2
itching
Mitracarpus villosus Cham. & Schlecht. PA2002/6 Dermatitis; wound; Leaf Infusion Topical 3
leprosy
Solanaceae Solanum verbascifolium L. PA2003/5 Dermatitis Leaf Poultice Topical 2
a Only plants with fidelity values of 2 or more were used in bacteriological tests.
b Fc, fidelity level (number of respondents citing the same plant).
 G.A. Pesewu et al. / Journal of Ethnopharmacology 116 (2008) 102–111 105

Table 2
Mean diameter of zones in agar plates where bacterial growth was inhibited by plant extracts placed in wells of 6 mm diameter
Plant species Mode of extraction Mean diameters (mm)a of inhibition zones in 7 bacterial isolatesb

MRSA I MRSA II MSSA S. p. E. c. P. a. P. v

Alchornea cordifolia Chloroform 10 10 12
Ethanol 16 20 21 12 13 10 20
Water 30 31 32 18 15 13 21
Blender 31 32 34 18 15 13 24

Cassia alata Chloroform

Ethanol 11 14 16 10 10 15
Water 16 18 20 14 12 18
Blender 16 18 20 14 12 18
Cassia occidentalis Chloroform
Ethanol 20 22 25 18 11
Water 10 10 14 10 8 12
Blender 10 12 16 10 8 14

Cryptolepis sanguinolenta Chloroform 14 14 22 12 8 10

Ethanol 24 26 28 20 13 22
Water 18 22 24 16 10 20
Blender 22 25 28 18 10 22
Elaeophorbia drupifera Chloroform
Ethanol 19 20 24 18
Water 15 17 20 12
Blender 15 17 20 12
Gardenia ternifolia Chloroform
Ethanol 10 10 12 8
Water 12 14 18 10
Blender 12 14 18 10

Mitracarpus villosus Chloroform

Ethanol 25 25 28
Water 20 22 26
Blender 20 22 26

Ocimum viride Chloroform

Ethanol 12
Water
Blender
Persea americana Chloroform
Ethanol 26 28 28 21
Water 22 25 26 16 10 8 20
Blender 23 26 30 18 10 8 20

Psidium guajava Chloroform 16 18 20

Ethanol 20 22 25 20 15 10 18
Water 18 21 24 10 10 8 10
Blender 22 24 27 15 10 8 10
Rauwolfia vomitoria Chloroform
Ethanol 10 12 16
Water
Blender

Solanum verbascifolium Chloroform

Ethanol
Water 20 20 21
Blender 20 22 24
Vernonia amygdalina Chloroform 8 8 12
Ethanol
Water 10 10 14 8 10 12
Blender 10 10 14 8 10 12

Controls:
Allicin 100 ␮l of 0.5 mg ml−1 Water extract 32 35 35 22 10 14 20
Gentamicin (10 ␮g) Paper disc 25 25 25 20 16 23 18
Vancomycin (30 ␮g) Paper disc 20 22 24 21 NTc NT NT
Extracts were obtained as serial extractions in chloroform, ethanol and water, or in aqueous solution in a ‘Stomacher’ blender. Blanks indicate no detectable inhibition of bacterial growth.
a Values each represent the means of three replicates; the pooled S.D. of all means = 0.5 mm (all used 100 ␮l of 50 mg ml−1 solution).
b Bacterial isolates: MRSA I, UELSHB 102; MRSA II, UELSHB 103; MSSA, NCTC 6571; S. p., Streptococcus pyogenes UELSHB 333.; E. c., Escherichia coli ATCC 25922; P. a.,
Pseudomonas aeruginosa ATCC 27853; P. v., Proteus vulgaris UELSHB 241.
c Not tested.
106 G.A. Pesewu et al. / Journal of Ethnopharmacology 116 (2008) 102–111

0.048–50 mg ml−1 except for allicin which was tested in the
range 0.024–25 mg ml−1 .
3. Results
3.1. Ethnobotanical survey
The study identified 25 plant species with fidelity levels of
two or more used by herbalists to treat appropriate skin infec-
tions and conditions (Table 1). Herbalists used the leaves of
plants as sources of extracts in 60% of species, the roots in
16%, the stem-bark in 12% and the seeds, fruits or shoots in
4% each. Local herbal practitioners used a number of methods
to prepare these medicines some materials were prepared using
more than one method. These methods were, infusions or decoc-
tions of fresh or dried material (18 species), juice squeezed from
leaves (5 species), poultices prepared from pulped plant tissues
(3 species), alcoholic extraction (1 species), oil extraction (1
species) or latex (1 species). Medicines from Cassia alata, Oci-
mum viride and Rauwolfia vomitoria are prepared by more than
one method. All of the prescriptions are used topically and 16%
used both orally and topically (Table 1). The majority (76%) of
the plant species were collected from the wild but 24%, mainly
fast growing species, are cultivated for easy access.
3.2. Plant extracts
Amongst those plant species with antibacterial activity (listed
in Table 2) the yields of materials extracted using different meth-
ods varied. The ranges of percentage dry weights isolated from
extracts were between 0.6% and 4% using chloroform, between
3.2% and 16% using ethanol and between 2.6% and 16.4% using
water. Chloroform extracts produced the lowest yield of mate-
rial and with the exception of Alchornea cordifolia and Cassia
occidentalis; yields were generally lower from water than from
ethanol. The yields obtained with the ‘Stomacher’ blender were
in the range 4.4% and 16.0% equal to or slightly greater than
those obtained in water extractions. The stomacher method was
also quicker to use, taking minutes as opposed to overnight
extraction.
3.3. Antibacterial activity
The antibacterial activities of the variously dissolved extracts
varied according to the species of bacteria tested. In 13 of the 25
plants tested at least one extract produced a zone of inhibition
greater than 10 mm against at least one species. In the remaining
12 plants, no antibacterial activity was detected (Table 2). Zones
of inhibition produced from aqueous and ethanolic extracts

Table 3
Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) of extracts, obtained with the ‘Stomacher’ blender, of selected plant
species against seven bacterial isolates
Plant species MIC/MBC (mg ml−1 ) MRSA Ia MRSA II MSSA S. p. E. c. P. a. P. v.

Alchornea cordifolia MIC 3.1 1.6 0.4 6.3 6.3 12.5 1.6
MBC 12.5 6.3 0.8 25 50 50 6.3
Cassia alata MIC >50 25 6.3 50 50 12.5
MBC >50 >50 50 >50 >50 50
Cassia occidentalis MIC >50 >50 50 >50 >50 50
MBC >50 >50 25 >50 >50 >50
Cryptolepis sanguinolenta MIC 25 25 3.1 50 50 3.1
MBC 50 25 12.5 50 >50 12.5
Elaeophorbia drupifera MIC >50 >50 6.3 >50
MBC >50 >50 >50 >50
Gardenia ternifolia MIC >50 >50 12.5 50
MBC >50 >50 50 >50
Mitracarpus villosus MIC 12.5 6.3 6.3
MBC 50 25 6.3
Persea americana MIC 6.3 3.1 1.6 12.5 >50 >50 12.5
MBC 25 25 6.3 50 >50 >50 50
Psidium guajava MIC 12.5 12.5 3.1 25 >50 >50 50
MBC 50 50 12.5 50 >50 >50 >50
Solanum verbascifolium MIC 25 6.3 3.1 >50
MBC >50 >50 6.3 >50
Vernonia amygdalina MIC >50 >50 >50 >50 >50 12.5
MBC >50 >50 >50 >50 >50 >50
Controls:
Allicin MIC 0.1 0.1 0.1 0.1 0.2 0.2 0.1
MBC 0.2 0.2 0.1 0.2 >0.2 >0.2 0.2
Gentamicin MICa 0.2 0.2 0.2 0.4 0.8 0.4 0.8
MBCa 0.8 0.8 0.8 3.1 3.1 3.1 3.1
Vancomycin MICa 0.8 0.8 0.4 0.8 NTc NT NT
MBCa 3.1 1.6 1.6 3.1 NT NT NT

Blanks indicate no inhibition. a Values are in mg l−1 ; b Abbreviations: MRSA I, UELSHB 102; MRSA II, UELSHB 103; MSSA, NCTC 6571; S. p., Streptococcus
pyogenes UELSHB 333; E. c., Escherichia coli ATCC 25922; P. a., Pseudomonas aeruginosa ATCC 27853; P. v., Proteus vulgaris UELSHB 241; c NT, not tested.
 G.A. Pesewu et al. / Journal of Ethnopharmacology 116 (2008) 102–111 107

Table 4
Mean diameter of zones where growth of MRSA isolates was inhibited by plant extracts

Plant species Mean diameters (mm)a of inhibition zones for 15 MRSA isolatesb

A1 A2 A3 A4 A5 B1 B2 B3 C1 C2 D E F G 102

Alchornea cordifolia 26 25 30 29 28 20 15 28 30 27 27 30 21 28 31
Cassia alata 10 15 14 20 15 16
Cassia occidentalis 12 12 8 10 14 18
Cryptolepis sanguinolenta 24 22 24 22 20 22 29 23 26 30 10 26 22
Elaeophorbia drupifera 10 8 10 15 12 14
Gardenia ternifolia 12 16 12 12 22 14 15 20 10 10 20
Mitracarpus villosus 15 18 18 21 22 20
Persea americana 16 20 18 10 20 17 18 19 10 24
Psidium guajava 20 22 20 21 20 20 21 20 20 24 18 23
Solanum verbascifolium 10 11 10 10 12 10 20
Vernonia amygdalina 8 12 8 10 10
Controls:
Allicin
100 ␮l of 0.5 mg ml−1 30 32 34 32 32 30 24 32 44 32 28 30 25 31 32
Gentamicin 10 ␮g 26 25 25 25 26 26 18 25 40 24 24 24 22 21 25
Vancomycin 30 ␮g 17 20 21 20 20 21 16 20 30 20 21 20 20 18 20
a Values each represent the means of three replicates; the pooled S.D. of all means = 0.3 mm. (All used 100 ␮l of 50 mg ml−1 solution).
b MRSA isolates and their origins: A: 1, 2, 3, 4, 5 – nose; B: 1, 2, 3 – leg ulcer; C: 1, 2 – wound swab; D: stoma; E: penis; F: vagina; G: eye; 102: UELSHB 102
(control).

and were generally similar but antibacterial activity was found
only in the ethanolic extracts of Ocimum viride and Rauwolfia
vomitoria and only in the aqueous extracts of Vernonia amyg-
dalina. In chloroform extracts antibacterial activity was found
in only four plant species and the inhibition zones were gen-
erally smaller than those from ethanolic or aqueous extracts.
The aqueous extracts of Alchornea cordifolia, Persea ameri-
cana and Psidium guajava inhibited the growth of all bacterial
species tested. Those of Cassia alata, Cassia occidentalis,
Cryptolepis sanguinolenta, Elaeophorbia drupifera, Gardenia
ternifolia, Mitracarpus villosus, Solanum verbascifolium and
Vernonia amygdalina inhibited the growth of fewer bacteria but
were active against all three isolates of Staphylococcus aureus.
The most active extract of these was the ethanol extract from
Cryptolepis sanguinolenta, average zone size 19 mm (range, no
zone to 28 mm). No inhibitory zone was found against Pseu-
domonas aeruginosa. Only three species showed any activity
against Pseudomonas aeruginosa, Alchornea cordifolia aque-
ous extracts were most active, giving a zone size of 13 mm, this
compared well with the allicin control (14 mm). The diameters
of the inhibition zones of the aqueous extracts of Alchornea
cordifolia were similar to, or greater than those of all other plant
species against corresponding bacteria average 23 mm (range
13–32 mm) (Table 2). In addition, the blender aqueous extracts
of Alchornea cordifolia showed the greatest activity, average
zone size 24 mm (range 13–34 mm) especially against Staphy-
lococcus aureus. These results were also reflected in lower MICs
(range, 0.4–3.1 mg ml−1 ) and MBCs (range, 0.8–12.5 mg ml−1 )
than those of the other plant species (Table 3).
Blender extracts, from the 11 plant species showing antibac-
terial activity in aqueous extracts (Table 2) were tested against
a further 14 clinical isolates of MRSA plus UELSHB 102
(Table 4). The extract of Alchornea cordifolia inhibited the
growth of all 15 MRSA isolates and the diameters of the inhi-
bition zones (average 26 mm, range 15–31 mm) were similar
to, or greater than, those of the other plant species against cor-
responding bacteria. Compared with the activity of Alchornea
cordifolia extracts, MRSA were less inhibited by the extracts
from the other species tested (Table 4).
4. Discussion
Chloroform extracts generally showed lower activity than
ethanolic or aqueous extracts (Table 2) and they inhibited a
smaller range of bacteria. There was no other indication of
variation in the ranges of antibacterial activity in the different
extracts, which might suggest that the solvents had extracted dif-
ferent antimicrobial substances from the same plant. Although
the activity of ethanolic extracts was generally greater than those
of aqueous extracts, the use of an aqueous based ‘Stomacher’
Lab-Blender technique was preferred because it is quicker to
use, and more efficient than the alternative methods. Samples
were enclosed in a disposable plastic Stomacher bag and there
is no direct contact between the sample and the device. The
blender requires no cleaning between operations, two or more
sample bags can be processed together and the extraction time
(5 min) is shorter than for the various alternative methods used
here or reviewed by Ong (2004).
Antibacterial activity has previously been reported (Table 5)
in 10 of the 13 plant species listed in Table 2. In the present study,
the range of bacteria against which activity is attributed, is ampli-
fied and in addition aqueous extracts, often excluded in previous
studies, were tested. We also tested the activities of extracts
against clinical isolates of MRSA and in many cases high-
lighted the potential for these extracts to be used against such
multiply antibiotic resistant organisms. Voss and Doebbeling
(1995) pointed out that MRSA is a problem for the world and
some hospitals in Africa have reported an increase in MRSA
108 G.A. Pesewu et al. / Journal of Ethnopharmacology 116 (2008) 102–111
Table 5
Published data on antibacterial activity of plant extracts of species listed in Table 2
Plant species Bacteria against which activity has been demonstrated
Alchornea cordifolia Staphylococcus aureus, Bacillus subtilis, Escherichia coli,
Pseudomonas aeruginosa, Klebsiella pneumoniae
Cassia alata Bacillus spp., Lactobacillus casei, Stapylococcus spp.,
Streptococcus spp., agrobacterium tumefaciens, Citrobacter
freundii, Enterobacter aerogenes, Escherichia coli,
Klebsiella pneumoniae, Neisseria gonorrhoeae, Proteus
spp., Pseudomonas aeruginosa, Salmonella spp., Serria
marcescens, Trichomonas vaginalis
Cassia occidentalis Staphylococcus aureus, Bacillus subtilis, Escherichia coli,
K. aerogenes, Proteus vulgaris, Ps. aerogenes
Cryptolepis sanguinolenta Campylobacter coli, C. jejuni, Staphylococcus aureus,
Streptococcus faecalis, Pseudomonas aeruginosa,
Escherichia coli, Salmonella typhimurum, Shigella
dysenteriae, Vibro cholerae, Mycobacterium fortuitum
Gardenia ternifolia Campylobacter spp, Staphylococcus aureus
Mitracarpus villosus Staphylococcus aureus, Bacillus subtilis, Streptococcus
faecalis, Escherichia coli
Ocimum viride Staphylococcus aureus, Streptococcus faecalis,
Pseudomonas aeruginosa, Proteus vulgaris, Bacillus
subtilis, B. cerus, Escherichia coli, Enterococcus faecalis
Persea americana Streptococcus mutans
Psidium guajava Staphylococcus aureus, Mycobacterium phlei
Vernonia amygdalina Escherichia coli, Salmonella typhi, Shigella spp., Bacillus
spp., Klebsiella pneumoniae, Pseudomonas aeruginosa,
Proteus vulgaris, Serria marcescens, Streptococcus spp.,
MSSA, MRSA.
isolation from 2% in 1985 to 50% in 1987. It is the view of our-
selves and others (Hancock, 2007), that novel sources of agents
active against MRSA, and other drug resistant micro-organisms,
should be actively sought. This is the reason we highlighted the
activity of these extracts against MRSA in this study. In many
cases our findings agreed with, or showed greater antimicro-
bial activity, to that of earlier workers. In some cases however
our findings showed less activity. Such variations were possibly
due to the different extraction methods employed, the sources
of plant materials and/or the different strains of bacteria tested.
The fact that different extraction methods can affect antibacterial
activity has been reported in earlier studies (Eloff, 1998).
Aqueous extracts from Psidium gujava were particularly
active against all 7 test strains with zones sizes of 24–27 mm
for Staphylococcus aureus. The activity of aqueous extracts and
ethanol extracts were similar the exception being the relatively
higher activity of the ethanol extract against Proteus vulgaris
(18 mm zone compared to 10 mm from aqueous). Abdelrahim
and co-workers demonstrated that methanol extracts from the
bark of P. gujava were more active than aqueous extracts
against Escherichia coli. Their extracts were particularly rich
in tannins. Gram positive organisms, including Staphylococcus
aureus were not as susceptible. In keeping with the findings of
Abdelrahim et al. ( 2002), we found ethanol extracts more active
against Gram negative strains than aqueous extracts.
The extracts from Cassia occidentalis were active against
six of the seven original test strains; the greatest activity was
from the ethanol extracts. From an ethanolic extract of roots
of Cassia occidentalis Chukwujekwu et al. (2006) isolated the
Authors
Ajao et al. (1985), Okeke et al. (1999), Tona et al. (1999), Ajali
(2000), Ebi (2001)
Khan et al. (2001), Somchit et al. (2003)
Perez and Anesini (1994), Samy and Ignacimuthu (2000)
Paulo et al. (1994), Silva et al. (1996), Cimanga et al. (1996),
Cimanga et al. (1997), Gibbons et al. (2003), Sawer et al. (2005)
Silva et al. (1996)
Irobi and Daramola (1994), Okunade et al. (1999), Bisignano et al.
(2000)
Orafidiya et al. (2001), Ngassoum et al. (2003)
Ofek et al. (2003)
Jairaj et al. (1996), Nascimento et al. (2000), Abdelrahim et al.
(2002)
Akinpelu (1999), Iwalokun et al. (2003)
anthraquinone, emodin. They showed this agent was bacte-
riostatic against Gram positive organisms at very low levels
(0.0078 mg ml−1 ). Our crude ethanolic extracts from Cassia
occidentalis leaves also produced better activity against Gram
positives than aqueous extracts. We additionally found that the
aqueous extracts from the leaves were not only bacteriostatic but
also bactericidal against MSSA (at 25 mg ml−1 ) this gives the
possibility that at higher concentrations, Cassia-emodin could be
bactericidal. This could be expected as emodin from other plants
(e.g. Aloe-emodin) has been reported as bactericidal, affecting
N-acetyl tranferase activity in some bacteria, Helicobacter pylori
(Wang et al., 1998; Ferro et al., 2003).
In contrast to the generally broad-spectrum activity from the
plant extracts highlighted above, our extracts from the leaves
of Solanum verbascifolium only produced activity against the
three Gram positives. Glycosidal alkaloids from the fruits of
Solanum plants have been associated with antibacterial activ-
ity in the past (McKee, 1959; Beaman-Mbaya and Muhammed,
1976; Fukuhara and Kubo, 1991). Solanum crystals from berry
juice have strong activity against Gram positive bacteria and
in addition are potent antifungal agents. Our aqueous extract
from the leaves of Solanum verbascifolium showed particular
activity against MSSA and MRSA and was bactericidal against
MSSA (6.3 mg ml−1 ). There is some suggestion however, that
although steroidal alkaloids have been associated in the past with
antibacterial activity, that the antibacterial agents in the berry
juice crystals are different and may be a phosphorylated purine
and not steroidal alkaloids (Beaman-Mbaya and Muhammed,
1976).
 G.A. Pesewu et al. / Journal of Ethnopharmacology 116 (2008) 102–111
The antibacterial activities of extracts from Elaeophorbia
drupifera, Rauwolfia vomitoria and from the leaves of Solanum
verbascifolium are reported here for the first time. Activity
against MRSA was also reported for the first time for many of
these plant extracts. Overall the aqueous (Stomacher) extracts
from Alchornea cordifolia, Psidium guajava and Persea ameri-
cana were the most active antimicrobials found in this study.
Tona and co-workers previously reported that extracts made
from the root bark of Alchornea cordifolia demonstrated antibac-
terial, antiamoebic and antispasmodic action and as such had
the potential to act as an anti-diarrheic agents (Tona et al., 1998;
Tona et al., 1999). In our hands, aqueous (Stomacher) extracts
from leaves were more active against MRSA and MSSA than
against common intestinal bacterial pathogens. Against these
organisms the extract was mainly bacteriostatic. We showed that
an aqueous extract from Alchornea cordifolia was active against
all MRSA strains (Table 4) and bactericidal against Staphy-
lococcus aureus (Table 3). Although the MICs and MBCs of
Alchornea cordifolia extracts were higher than those of the con-
trols; purified allicin (Cutler and Wilson, 2004), gentamicin and
vancomycin, one has to remember that the Alchornea cordifolia
extract was a crude agent compared to these controls. When the
active antibacterial agent of Alchornea cordifolia is isolated and
purified its relative activity may increase. Ebi (2001) reported
the inhibition of bacterial growth by leaf extracts of Alchornea
cordifolia. Ebi found chloroform soluble extracts were active
against Staphylococcus aureus, Pseudomonas aeruginosa and
Escherichia coli but demonstrated greater anti-staphylococcal
and anti-pseudomonal activity in the fractions that were insol-
uble in chloroform. Okeke et al. (1999) also reported that leaf
extracts from Alchornea cordifolia were active against Gram
positives. The activity they reported was similar but slightly
lower than ours with MICs ranging from 5 to 20 mg ml−1 . We
have also shown that Alchornea cordifolia extracts are not only
inhibitory but can be bactericidal. Staphylococci were found to
be most susceptible with MICs below 3.1 mg ml−1 and MBCs
ranging for 0.8–12 mg ml−1 (concentrations that are 2–4 times
the MIC). As to current knowledge on the toxicology of aqueous
extracts of Alchornea cordifolia, a recent paper in a veterinary
journal identified Alchornea cordifolia as one of five plants of the
Euphorbiaceae family suspected of being poisonous to grazing
animals. They reported that giving a crude aqueous extract orally
to rats caused significant, potentially toxic, changes in blood
chemistry. They also suggested that the tannins in the extract
were responsible for these changes (Adedapo et al., 2007). In
contrast to this, tannins from the same plant have been proposed
as a possible adjuvant to penicillin in treating Staphylococcal
skin infections. It is thought that they reduce the ability of the
organism to coagulate plasma and form antibiotic resistant bac-
terial biofilms in wounds (Akiyama et al., 2001). Other workers
have also reported from tests on mice, that the crude ethanol
extract can be effective in treating Staphylococcus aureus infec-
tions at oral doses between 4 and 32 times lower than the lethal
dose (Igbeneghu et al., 2007). Given the interesting potential
of this agent against MRSA, further work is planned to sepa-
rate out the various phytochemicals in our aqueous extract and
test them for antimicrobial and toxicological activity. Current
109
evidence suggests the crude aqueous extract may be toxic if
taken orally however its use as a topical agent may provide a
potential way forward, as was suggested above by Akiyama and
co-workers.
In contrast to the active agents discussed above, some of
our extracts showed relatively low antibacterial activity but
our findings compared well to the work of other workers,
although different extraction methods and plant organs were
sometimes used. Examples of these were extracts from Ver-
nonia amygdalina and Cassia alata. Vernonia amygdalina is
a vegetable regularly consumed in large amounts by Nige-
rians and other West Africans. Rotimi et al. (1988) found
aqueous extracts of Vernonia amygdalina were very active
against oral anaerobic bacteria. Currently aqueous extracts
of Vernonia amygdalina are also under investigation as anti-
tumour agents. Studies into its cytotoxicity using MTT assays
showed no cytotoxic effects in the concentration range of
3–25 mg ml−1 . This was an improvement to the known toxic
effects from chloroform extracts. These contain vernodaline,
vernolide, and vernomygdine three agents reported to be toxic
to 50% of cells at approximately 2000 mg ml−1 (Izevbigie,
2003). We found chloroform extracts from Vernonia amyg-
dalina had very low antibacterial activity (<12 mm zones) but
our aqueous extracts produced slightly better activity (<14 mm
zones) against six of the original seven test strains. Iwalokun
et al. (2003) found that their aqueous extracts of Vernonia
amygdalina also low activity with zones of inhibition of approx-
imately 16 mm against MRSA and MIC’s of 22–26 mg ml−1 .
Our V. amygdaline extracts failed to produce such activity
against MRSA and had MIC’s of >50mg ml−1 . By compar-
ison, when the purified sesquiterpine lactones from Vernonia
amygdaline; vernolide and vernodalol were tested, Erasto et al.
(2006) reported MIC’s as low as 0.5 mg ml−1 against Staphy-
lococcus aureus. These purified compounds were not tested
against MRSA.
Cassia alata extracts have been reported to produce low lev-
els of activity against a wide range of micro-organisms (Khan
et al., 2001; Somchit et al., 2003). The greatest activity came
from petrol extracts of flowers with zones of 20 mm produced
against Staphylococcus epidermidis (Khan et al., 2001). Leaf
extracts produced no more than 16 mm zones of inhibition,
aqueous extracts were not tested. Somchit et al. (2002) found
water and ethanol extracts from barks produced a maximum
zone of 16 mm against Staphylococcus aureus and Escherichia
coli. These results were comparable to those found in our study;
in fact our aqueous extracts produced equal activity to the petrol
extracts cited above.
The present study has shown that medicinal plants, collected
following ethnobotanically directed interviews in Ghana; pos-
sess antibacterial properties that support their value in herbal
medicine for the treatment of diseases and aliments. These
studies highlight the importance in using an ethnomedicinal
approach to identifying plants from which new therapeutic
agents can be obtained. We specifically targeted topical anti-
infectives in this study but, since the diagnosis of skin infections
without the laboratory backup to determine the true cause of the
disease is difficult, agents were selected with a general history of
110 G.A. Pesewu et al. / Journal of Ethnopharmacology 116 (2008) 102–111
being used to treat skin complaints, as well as those recognized
as broad-spectrum antimicrobial agents. The one exception to
this was Cryptolepis sanguinolenta, this plant is used as a sys-
temic antimicrobial against malaria. In our study, we also found
that the extract was active against MRSA. Although there is no
ethno botanical background for the use of Cryptolepis sanguino-
lenta extracts as an anti-infective, other workers have previously
reported that Cryptolepis sanguinolenta alkaloids had antibac-
terial activity (Paulo et al., 1994; Cimanga et al., 1996) and the
work of Sawer et al. (2005) further demonstrated that the alka-
loid cryptolepine in particular had anti-staphylococcal activity
(against MSSA). We have additionally demonstrated that Cryp-
tolepis sanguinolenta extracts were active against a wide range
of MRSA and using culture methods, confirmed the electron
microscopy findings of Sawer that Cryptolepis sanguinolenta
extracts are bactericidal. In their paper cryptolepine was bacte-
ricidal at four times the MIC; this is similar to our findings for
MSSA. Another point of interest is that the extract showed lit-
tle activity against the Gram negative organisms, Escherichia
coli and Pseudomonas aeruginosa and was primarily active
as a Gram positive antimicrobial. One potential problem with
using cryptolepine extracts is the cytotoxicity. Earlier Cimanga
et al. (1996) reported cryptolepine to be “quite” cytotoxic.
This was also confirmed by Ansah and co-workers (Ansah and
Gooderham, 2002; Ansah et al., 2005). They compared aqueous
Cryptolepis sanguinolenta extracts with purified cryptolepine.
They found that the aqueous extract and the pure cryptolepine
were both cytotoxic and weak mammalian mutagens. How-
ever, they also suggested that because of the poor genotoxicity
of the compounds and their potent cytotoxic action that they
could be possible anticancer agents. This reported cytotoxic-
ity may however be a problem with its use as an antibacterial
agent.
The plant extracts demonstrating the highest levels of
antibacterial activity were from Alchornea cordifolia, Persea
americana and Psidium guajava. These all had an ethno botan-
ical history for treating skin complaints. Psidium guajava
had previously been reported to be antibacterial and active
against Staphylococcus aureus (Qadan et al., 2005). We have
for the first time demonstrated its activity against MRSA.
Persea americana extracts have only been reported as active
against Streptococcus mutans, reducing is adherence capabili-
ties (Ofek et al., 2003). We have now shown extracts additionally
possess antibacterial and more specifically anti-MRSA activ-
ity.
5. Conclusions
Multiple drug resistance in bacterial pathogens is a continu-
ing problem throughout the world. There is an established need
to develop new antimicrobial agents to combat these pathogens.
In our study, we found that 11 out of 13 crude plant extracts with
a history of ethnobotanical use in Ghana for the treatment of
wounds, contain antibacterial compounds that we have demon-
strated have a potential use against MRSA and other nosocomial
pathogens. Further work is ongoing to identify the exact nature
of these antimicrobials.
Acknowledgements
We thank the late Mr. Victor Biako and Mr. Ofori Lartey of the
Centre for Scientific Research into Plant Medicine (CSRPM),
Mampong-Akwapim, Ghana, for assistance in plant identifica-
tion and the preparation of herbarium voucher specimens, Dr. P.
Wilson of the Royal London Hospital, UK for supplying isolates
of MRSA, and Dr. T. Hill and Prof. A.V. Roberts (UELSHB) for
their help.
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