Assessment of Antimicrobial Activity of the Essential Oil from the Stem Powder of Cissus populnea and the Leaves of Sesamum radiatum, Herbal Medications for Male Infertility Factor
One of the causes of male infertility factor is venereal diseases. Cissus populnea and Sesamum radiatum are two tropical medicinal plants used to correct male infertility factor. The aim of this study was to extract the essential oil from the stem powder of C. populnea and the leaves of S. radiatum and screen them for antimicrobial activity. The essential oil from the two plants were obtained by hydro-distillation and were both found to inhibit the growth of Gram-negative and Gram-positive bacteria including Citrobacter sp., Escherichia coli, Escherichia coli ATCC 25922, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus albus and Staphylococcus aureus. The essential oil from C. populnea was in addition found to be active against Enterococcus faecalis, Staphylococcus aureus ATCC 25923 while the essential oil from S. radiatum was found to be active against Serratia marcescens. The essential oil samples from the two plants were both inactive against Candida albicans. The results confirm the antimicrobial properties of the essential oil from the two plants and hence, these plants may correct male infertility factor arising from bacterial infection.
to cite this article:
E.A.S. Osibote, M. Ogunlesi, W. Okiei, T. Asekun and O.B. Familoni, 2010. Assessment of Antimicrobial Activity of the Essential Oil from the Stem Powder of Cissus populnea and the Leaves of Sesamum radiatum, Herbal Medications for Male Infertility Factor. Research Journal of Medicinal Plants, 4: 14-20.
Cissus populnea Guill and Per belongs to the Vitaceae family. It is
used extensively in medicinal preparations in West Africa. In Benin Republic,
it is used as a diuretic and in Ghana it is used as a post-harvest ethnobotanical
protectant (Belmain et al., 2000). The aqueous
extract of the stem bark is used as a fertility enhancer in males in South Western
Nigeria (Ojekale et al., 2006). The stem bark
has been shown to contain tannins, flavonoids, saponins and steroids (Ojekale
et al., 2006). Extracts from the root of the plant have been used
for the management of skin diseases, boils, infected wounds (Kone
et al., 2004) and for treating urinary tract infections (Ojekale
et al., 2006) thus suggesting antibacterial activity. Anthraquinone
derivatives, steroidal and cardiac glycosides have been reported present in
the root extract while alkaloids and tannins were found to be absent (Moody
et al., 2003).
Sesamum radiatum Schum. and Thonn belongs to the Pedaliaceae family
(Hutchinson and Dalziel, 1954). The decoction of the leaves
is used for the treatment of catarrh, eye pains, bruises and erupted skins (Bankole
et al., 2007). The decoction of combined roots and leaves has been
reported to exhibit anti-viral and anti-fungal activity (Gill,
1992). The aqueous extract of the leaves has been reported to be rich in
phenols, flavonoids, lignans and sterols (Shittu et al.,
2006). The phenols would be expected to exhibit antimicrobial activity (Konan
et al., 2008).
The two plants investigated are used in treating male infertility factor in
South-Western Nigeria. It has been reported that venereal diseases could be
responsible for male infertility factor (Greendale et
al., 1993). It is thus possible that these plants may exhibit antimicrobial
activity which may cure venereal diseases and thus correct male infertility
factor arising from bacterial infection. This is the reason for this investigation
of the antimicrobial activity of the essential oils from these plants.
MATERIALS AND METHODS
Collection of Plant Materials
Several batches of the fresh stem of C. populnea and fresh leaves
of S. radiatum were obtained from Mushin market in Lagos between the
months of October 2005 and May 2007. They were identified and authenticated
by Mr. T.K. Odewo and Mr. Seun Osiyemi both of the Forestry Research Institute
of Nigeria (FRIN), Ibadan. A voucher specimen for C. populnea with the
number FHI 108222 was deposited at the Herbarium in FRIN on 27th May, 2008 while
the voucher specimen for S. radiatum with number FHI 107823 was deposited
at the Herbarium in FRIN on 12th September, 2007.
Hydro-Distillation of Samples
The plant parts were cut into small pieces and air-dried at room temperature
in a dust-free environment for 3 weeks. The dried materials were powdered and
the essential oil from each plant was extracted by hydro-distillation in batches
of 100 g mixed with 3 L of water. The essential oil was extracted into hexane
and the solution obtained was concentrated by evaporation of the hexane. The
volume of the essential oil was 0.3 cm3 and weighed 350 mg.
Preparation of Test Organisms
The test microorganisms used were collected from the stock cultures of the
Medical Microbiology and Parasitology Department of the College of Medicine,
University of Lagos, Idi-Araba, Surulere, Lagos. They included Candida albicans
(a yeast) and Gram-negative and Gram-positive bacteria, namely Citrobacter sp.,
Enterococcus faecalis, Escherichia coli, Escherichia coli ATCC25922,
Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa,
Serratia marcescens, Staphylococcus albus, Staphylococcus aureus
and Staphylococcus aureus ATCC 25923.
The essential oil samples from the two plants were screened for antimicrobial
activities using the agar disc diffusion method (Dosso and
Kette, 1995). The test organisms were subcultured on Blood Agar and Mac-Conkey
Agar (Oxoid, UK). Suspensions of the microorganisms were prepared in sterile
normal saline and adjusted to 0.5 McFarland standards. The resulting suspension
contained approximately 1x108 cfu cm-3. Each medium plate
was uniformly seeded with a sensitive strain of each microorganism. The sample
of each essential oil was dissolved in hexane to give a concentration of 260
μg cm-3 and sterile paper discs, 6 mm diameter, were soaked
in each solution. The discs were transferred to the inoculated plates immediately.
After refrigeration at 4°C for 1 h, the plates were incubated at 37°C
for 24 h. The 0.05% ciprofloxacin in hexane and neat hexane were used as controls.
Zones of inhibition around the discs were measured in millimeter and used as
an assessment of antimicrobial activity.
results of antimicrobial screening of the samples of essential oil obtained
from the stem of Cissus populnea and the leaves of Sesamum radiatum
5-9 mm zone of inhibition; 2+: 10-19 mm zone of inhibition; 3+: > 20
mm zone of inhibition
The results of antimicrobial screening of the samples of essential oil obtained from the stem of C. populnea and the leaves of S. radiatum are shown in Table 1. Hexane did not inhibit the growth of any of the microorganisms. The essential oil obtained from the stem of C. populnea inhibited the growth of ten microorganisms. It strongly inhibited E. coli, P. aeruginosa and S. aureus, exhibiting the same extent of inhibitory zone of 3+ as the control, 0.05% ciprofloxacin. There was significant but lower activity with inhibitory zone of 2+ against Citrobacter sp., E. faecalis, E. coli ATCC 25922, K. pneumoniae, P. mirabilis, S. albus and S. aureus ATCC 25923. There was no activity against C. albicans and S. marcescens.
The essential oil from the leaves of S. radiatum inhibited the growth of nine microorganisms, with no activity against C. albicans, E. faecalis and S. aureus ATCC 25923. The activity against the other microorganisms namely Citrobacter sp., both strains of E. coli, K. pneumoniae, P. mirabilis, P. aeruginosa, S. marcescens, S. albus and S. aureus all showed inhibitory zone of 2+. The essential oil from S. radiatum was consistently less potent than the ciprofloxacin control.
In comparison, the essential oil obtained from C. populnea inhibited all the Gram-positive microorganisms tested namely E. faecalis and the three staphylococcus strains while the essential oil from the leaves of S. radiatum inhibited the growth of two Gram-positive bacteria tested namely S. albus and S. aureus, showing no inhibitory action against S. aureus ATCC 25923 and E. faecalis. The essential oil from C. populnea exhibited higher activity against E. coli, P. aeruginosa, both strains of S. aureus and E. faecalis but no activity against S. marcescens compared to the sample from S. radiatum.
The results show that the samples of the essential oil from the two plants
have great potential as antimicrobials hence might be able to correct male infertility
factor arising from infection by the bacterial species. Extracts obtained from
C. populnea have been reported to be useful for treating skin diseases,
boils (Kone et al., 2004) and urinary tract infections
(Ojekale et al., 2006).
In a screening of 50 medicinal plants used in North Cote-dIvoire as traditional
remedies for bacterial diseases, C. populnea was among the ten plant
species which exhibited promising level of activity against bacteria including
strains resistant to aminosides, macrolides, penicillin M, lincosamide and streptrogramin
B (Kone et al., 2004). The ethanolic root extract
of C. populnea was used in the study. The bacteria used in the study
included Gram-positive strains, S. aureus, E. faecalis, B.
subtilis and Streptococcus pyogenes as well as Gram-negative strains
such as E. coli and P. aeruginosa. The root extract was not active
against E. coli and P. aeruginosa but was found to be active against
a methicillin-resistant hospital strain of S. aureus and some aminoside-resistant
hospital strains of E. faecalis. In our study, the essential oil of
C. populnea was found to be significantly active against E. faecalis,
the three staphylococcus strains and even against E. coli and P. aeruginosa
which were not inhibited by the root extract (Kone et
In a study on the antimicrobial activity of ethanolic and aqueous extracts
of the dried leaves of S. radiatum against five microorganisms, namely
S. pneumoniae, C. albicans, S. aureus, E. coli
and P. aeruginosa, it was observed that the ethanolic extract exhibited
mild inhibitory effects on S. pneumoniae and C. albicans while
the aqueous extract of the same concentration did not show any inhibitory effects
(Shittu et al., 2006). The essential oil in our
study did not exhibit activity against C. albicans but was active against
S. aureus, E. coli and P. aeruginosa.
The aqueous extract (Shittu et al., 2006) was
obtained by boiling the aqueous suspension of the powdered leaves for 3 h and
thus some of the constituents of essential oil in the leaves would have been
extracted and thus would be expected to show some inhibitory effects against
some of the microorganisms tested as was observed in the study being presented
in our report. However, the aqueous extract was lyophilized and this process
might have resulted in extensive loss of the essential oil constituents. This
may explain the inactivity of the aqueous extract.
Several medicinal plants which are used for the management of male infertility
factor in Nigeria are also used as antimicrobials. Such plants include Abrus
precatorius, Adenia venata, Agelenopsis chevaleri, Aloe
barteri, Areca catechu, Craterispermum laurinum, Circuligo
pilosa, Datura stramonum, Ritchiea brachypoda, Saba florida,
Securidaca longepedunculata, Tabernaemontana pachysiphen, Tribulus
terrestris and Withania somnifera several of which are used for treating
venereal diseases and urinary tract infections (Odugbemi,
2008a). Mucuna pruriens which is also used as a male fertility enhancer
is used as a phytomedicine for genito-urinary diseases (Odugbemi,
2008b). Some phytomedicines classified as aphrodisiacs are alleged to be
antimicrobials in herbal preparations. In a review on Infections and male infertility,
Megory et al. (1987) stated that bacteria could
affect semen quality and cause changes which could impair fertility function.
Infections could cause inflammation of the accessory gland or total damage to
the gonads and adrexa (Megory et al., 1987).
Greendale et al. (1993) reported that venereal
diseases could reduce fertility in male subjects. Thus the use of antimicrobials
is relevant in the management of male infertility.
The essential oil from Syzygium aromaticum (L.) Merr. and Perry, a herbal
aphrodisiac has been reported to exhibit significant antimicrobial activity
against a collection of 25 different genera of test bacteria and 20 different
isolates of Listeria monocytogenes (Deans et
al., 2006). It was also reported to exhibit high levels of inhibition
against three fungal strains of a plant pathogen (Deans
et al., 2006). Terminalia catappa another herbal aphrodisiac
has also been reported to show promising antibacterial properties. In the antimicrobial
screening of nine Peruvian medicinal plants, the leaves of Terminalia catappa
and aerial parts of Phyllantus amarus showed the most promising antibacterial
properties inhibiting all the bacterial strains tested with minimum inhibitory
concentrations ranging from 0.25 to 16 mg cm-3 (Kloucek
et al., 2005). The microorganisms used were Bacillus cereus
ATCC 11778, B. subtilis ATCC 6633, Bacteroides fragilis ATCC 25285,
E. faecalis ATCC 2912, E. coli 25922, P. aeruginosa ATCC
27853, S. aureus ATCC 25923, S. epidermidis ATCC 12228 and Streptococcus
pyogenes ATCC 19615. The ethanolic extracts of T. catappa demonstrated
activity in the following decreasing order: S. epidermidis ATCC 12228
> S. aureus 25923 > B. cereus > B. subtilis
= P. aeruginosa > E. coli 25922 = E. faecalis
> S. pyogenes ATCC 19615 = B. fragilis. In our study the essential
oils from the two plants were active against the two strains of E. coli
one of which was strain ATCC 25922 as well as P. aeruginosa. In addition,
the essential oil from C. populnea was active against E. faecalis
and S. aureus. Thus these results are similar to those on T. catappa.
High antifungal but no antibacterial activity of methanol and methylene chloride
extracts from T. catappa aerial part was observed (Goun
et al., 2003). However, Pawar and Pal (2002)
detected appreciable antimicrobial activity of the chloroform and methanol
extracts of the roots of T. catappa against E. coli and S.
Tribulus terrestris, a herbal aphrodisiac, analgesic, diuretic, anti-hypertensive
agent also used for the management of urinary disorders, impotence and gonorrhea
was investigated for antibacterial activity (Kianbakht and
Jahaniani, 2003). The methanolic extracts of Iranian plant parts, fruits,
stems, leaves and roots, showed considerable activity against all the bacteria
used in the study, namely S. aureus, E. faecalis, E. coli
and P. aeruginosa. However, the ethanolic extracts of Yemini plant
did not exhibit antibacterial activity against any of the reference bacteria
(Ali et al., 2001). However, some researchers
reported that all plant parts of the Turkish T. terrestris showed activity
against several reference bacteria and some researchers and noted that the ethanolic
extracts of the fruits and leaves of Indian T. terrestris were active
against E. coli and S. aureus (Kianbakht and
The reports on the studies on T. terrestris support antibacterial
activity in plant parts (except in the Yemeni plant) and hence its usefulness
in the management of urinary tract infection especially the activity against
E. coli, a microorganism which has been implicated in urinary
tract infection (Tena et al., 2008).
The report of our study on eleven bacterial samples and the bioactivity of the essential oils from the two plants on several of these microorganisms show that these essential oils are broad-spectrum antibacterial agents which may be useful in inhibiting the growth of some microorganisms which can be causative agents of infertility in male subjects.
The essential oil from the stem of C. populnea and the leaves of S. radiatum have been shown to demonstrate significant antibacterial activity against several bacterial strains including both Gram-positive and Gram-negative microorganisms.
The authors express appreciation to Prof. T. Odugbemi of the Department of Medical Microbiology and Parasitology of the College of Medicine, University of Lagos, for providing the microorganisms and the laboratory facility for the antimicrobial screening and Mrs. T. Ogunsanya for demonstrating the antimicrobial assay.
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