Content uploaded by Latifou Lagnika
Author content
All content in this area was uploaded by Latifou Lagnika on Mar 27, 2014
Content may be subject to copyright.
Indian Journal of Science • Analysis
Lagnika et al.
In vitro antibacterial activity of two medicinal plants used in Bénin to treat microbial infections,
Indian Journal of Science, 2014, 8(19), 10-15, www.discovery.org.in
http://www.discovery.org.in/ijs.htm © 2014 Discovery Publication. All Rights Reserved
Page
10
Lagnika L1☼, Amoussa AMO2, Sanni A3
1. Lecturer, researcher, Dept. of Biochem and Cell. Biol., Laboratory of Biochemistry and bioactive natural substances, University of Abomey-Calavi, 04 BP 0320,
Cotonou, Bénin, West Africa
2. PhD, Dept. of Biochem and Cell. Biol., Lab. of Biochem. and bio. Nat. Subst., Univ. of Abomey-Calavi, 04 BP 0320, Cotonou, Bénin, West Africa
3. Senior Lecturer, Dept. of Biochem and Cell. Biol., Lab. of Biochem. and bio. Nat. Subst., Univ. of Abomey-Calavi, 04 BP 0320, Cotonou, Bénin, West Africa
☼Corresponding author: Lecturer, researcher, Univ. of Abomey-Calavi, 04 BP 0320, Cotonou, Bénin, West Africa, Mail: latifkabe@yahoo.fr;
Mobile No: (+229) 97604889
Publication History
Received: 06 December 2013
Accepted: 23 January 2014
Published: 5 February 2014
Citation
Lagnika L, Amoussa AMO, Sanni A. In vitro antibacterial activity of two medicinal plants used in Bénin to treat microbial infections. Indian
Journal of Science, 2014, 8(19), 10-15
ABSTRACT
Dichloromethane, methanol and hydroethanol extracts of Casuarina equisetifolia L. and Oxalis corniculata L., two medicinal plants
used by traditional doctors to treat microbial infections, were screened for their antibacterial activity against seven Gram positive
and Gram negative bacteria (Escherichia coli, Staphylococus aureus, Enterococus faecalis, Pseudomona aeruginosa, Salmonella
aboni, Staphylococus aureus meticilline resisting and Staphylococus epidermidis). The antibacterial activity was performed using the
p-iodonitrotetrazolium microdilution method. The total activity and Artemia salina lethality of extracts were also determined. All
extracts were effective against tested microorganisms at different levels with Minimum Inhibitory Concentration values ranging
from 0.078 mg/ml to 5 mg/ml. The hydroethanolic extract was more potent than other extracts with a MIC value of 0.078mg/ml
against S. epidermidis. The most interesting total activity was obtained with hydroethanolic extract of Oxalis corniculata (1689.7
ml). The methanol extract of Oxalis corniculata was also the less toxic to Artemia salina with LC50 value of 26.87 mg/ml.
Keywords: Medicinal plants, antibacterial, toxicity, artemia salina
1. INTRODUCTION
Infectious diseases are becoming a crisis as a major cause of human and animal mortality and morbidity. This is
further aggravated by the rapid development of multi-drug resistance, limited antimicrobial spectrum and adverse
RESEARCH • MEDICINAL PLANTS Indian Journal of Science, Volume 8, Number 19, February 5, 2014
In vitro antibacterial activity of two medicinal plants used in Bénin to treat
microbial infections
S
ci
e
nce
Indian Journal of
ISSN 2319
–
77
30
EISSN 2319
–
77
49
Lagnika et al.
In vitro antibacterial activity of two medicinal plants used in Bénin to treat microbial infections,
Indian Journal of Science, 2014, 8(19), 10-15, www.discovery.org.in
http://www.discovery.org.in/ijs.htm © 2014 Discovery Publication. All Rights Reserved
Page
11
effects of available anti-microbial agents (Doughart and Okafor, 2007). In most countries of West Africa, the advent of
modern medicine and its advances have led people to shy away some traditional medicine, based primarily on herbal
medicine. Unfortunately, development of antibacterial agents has been accompanied by the emergence of drug-
resistant organisms followed by toxicity observed during prolonged treatment. The use of plants for healing dates
from prehistoric times and all peoples have this old tradition. The use of herbs in the treatment of man and animal
disease has been also practiced before the advent of modern antibiotics (George, 1974; Soforowa, 1982). Thus, in
recent decades, the use of medicinal plants has been renewed interest. Special attention is given to the search for
new and effective pharmaceutical agents, with little or no toxicity, from medicinal plants. Natural products and
related structures are essential sources of new pharmaceuticals, because of the immense variety of functionally
relevant secondary metabolites of microbial and plant species (Ngo et al. 2013). Approximately half of all drugs that
were recorded worldwide in the period before 2007 were from natural products or their synthetic derivatives
(Kennedy and Wightman, 2011). Several scientific studies have confirmed the activities of most of the plants used in
traditional medicine not only against microbial infections (Traoré et al. 2012; Bolou et al. 2011; Adedapo et al. 2009).
In Benin, plants materials have been used as traditional medicines for the treatment of a wide variety of ailments and
diseases. Casuarina equisetifolia and Oxalis corniculata are two plants commonly used in Benin traditionally medicine
against infections of the skin, dizziness, diarrhea, stomach ache, dysentery, convulsions and other digestive problems.
2. SCOPE OF THE STUDY
The present work is to investigate the antibacterial properties of Casuarina equisetifolia and Oxalis corniculata against
six strains of bacteria Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus epidermidis,
Staphylococcus aureus and Staphylococus aureus meticilline resisting. The lethality effect of extracts was also
evaluated using Artemia salina.
2.1. Materials
The materials used include fruits of Casuarina equisetifolia, leaves of Oxalis corniculata and others which include,
pipette, bottles, weighting balance, conical flasks, petri dishes, and electric blender, rotary evaporator, hood, oven,
mixer etc.
2.1.1. Reagent/chemical/median
p-iodonitrotetrazolium (Sigma Aldrich) solution was prepared by dissolving 4 mg in 200 ml distilled water. The Muller
Hinton broth (DIFCO) was prepared by dissolving 21 g in 1 L distilled water. The suspension was then sterilized using
autoclave.
2.1.2. Organisms used
Escherichia coli (CIP53126), Staphylococus aureus (ATCC6538), Enterococus faecalis (ATCC29212), Pseudomona
aeruginosa (CIP82118), Salmonella aboni (CIP8039), Staphylococus aureus meticilline resisting (SARM) and
Staphylococus epidermidis, obtained from Laboratoire de Biophotonique et Pharmacologie, University of Strasbourg,
France, were used for antibacterial activity. Eggs of Artemia salina were hatched in seawater to evaluate extracts
toxicity.
2.2. Methodology
2.2.1. Preparation of extracts
One hundred grams (100 g) of dry powder of each part of the two plants were successively extracted by maceration
with dichloromethane and methanol for 72 h stirring. A second maceration with a mixture of ethanol-water (20:80)
was carried out with fifty grams (50 g) of dry powder species. Each extraction is repeated three times. The macerates
were filtered and concentrated using a rotary evaporator (RE 300, stuart) and the extracts were stored at 4° C until
biological assay.
2.2.2. Antibacterial activity
Growth inhibitory effect of extracts: This test aims to eliminate the extracts which at 10 mg/ml do not inhibit the
growth of bacteria (Eloff, 1998). The extracts are prepared to a concentration of 20 mg/ml in a mixture
acetone/Mueller Hinton broth (v/v). The obtained solutions are then homogenized using vortex. 100 µl of each
extract at 20 mg/ml were distributed in triplicate wells of a 96 well plate containing previously 100 µl of bacterial
broth at 106 CFU/ml. The plates were homogenized using a mixer and then incubated at 37°C. After 18 h of
incubation, 40 µl of a 0.2 % of p-iodonitrotetrazolium (INT) (Sigma Aldrich) in distilled water, were added to each well
Comparison: This is
to check phased
manner, the
accuracy of the
data related to the
use of a product,
using methods
approved by
science. This audit
will confirm or
refute empirical
data taken by a
segment of the
population, based
on the results.
Content:
Verification of data
and past practices.
This will be
recorded in
documents such as
articles, books, etc.,
or disseminated
through the press.
Iodonitrotetrazolium:
It is an artificial electron
acceptor which can be
used in a colorimetric
assay to determine the
presence of protein in
culture median. The
presence of protein
such as threonine
desydrogenase during
active growth of
bacteria changes the
color of INT (incolore)
to purple-red.
Muller Hinton:
It is a microbiological
growth median that is
commonly used for
antibiotic, extracts or
natural compounds
susceptibility testing.
Lagnika et al.
In vitro antibacterial activity of two medicinal plants used in Bénin to treat microbial infections,
Indian Journal of Science, 2014, 8(19), 10-15, www.discovery.org.in
http://www.discovery.org.in/ijs.htm © 2014 Discovery Publication. All Rights Reserved
Page
12
and the plates were incubated again at
37°C. After 30 min of incubation, the color
change of the contents of the wells
(extract color to red) indicates bacterial
growth in wells and led to eliminate non-
active extracts. The Wells which
maintained their color after the addition
of INT contains active extracts.
Minimum Inhibitory Concentration (MIC): The Minimum Inhibitory Concentrations (MICs) were determined by the
method of broth microdilution using p-iodonitrotetrazolium (INT) as an indicator of bacterial viability (Keymanesh et
al. 2009). To determine the MIC of extracts, 100 µl of Mueller Hinton broth (DIFCO) were added to each wells of a 96-
wells microplate and then 100 µl of plant extract (20 mg/ml) were added to the first well (A) of the plate. A two-fold
Successive dilution was carried from well (A) to the last wells (H) of the plate. Then, 100µl of bacterial broth at 106
CFU/ml were finally added into all the wells. The plate was covered and incubated at 37°C. After 18 h of incubation,
40 µl of p-iodonitrotétrazolium (0.2 %) was added in all wells and the plates were incubated again at 37°C. After 1 h of
incubation, wells were examined and the MIC values were recorded.
Total activity: To select the extracts that can be used for further testing, the determination of the total activity is
important because since the MIC value is inversely proportional to the amount of antimicrobial extracts. The total
activity of each extract was calculated by dividing the MICs with the amount of extract obtained from 1 g of plant
material (Eloff, 2008). This value indicates the volume in which the active principle (extract) present in 1 g of dry plant
material can be diluted to always have inhibitory activity against organisms (Eloff, 2004).
2.2.3. Brine Shrimp Lethality Bioassay
The assay was performed as described by Keymanesh et al. (2009). To obtain mature naupli larva, the eggs of brine
shrimp were hatched in normal seawater for 72 h. The stock solution of each extract (3 mg/ml) was obtained by
dissolving 15 mg in 200 μL ethanol and 4.80 ml of seawater. Then, 1 ml of seawater containing 15 living naupli was
added to 1 ml of extracts. Six concentrations ranging from 1.5 to 0.075 mg/ml, obtained by a twofold dilution of stock
solution, were tested. Each experiment was done in triplicate and control was prepared using only seawater plus 15
livings naupli. Survivors naupli were counted after 24 h and dead naupli at each concentration were determined. The
percentage of lethality of the brine shrimp was then calculated.
3. RESULTS
The Dichloromethane, methanol and hydroalcoholic extracts of two traditional Beninese pharmacopoeias, Oxalis
corniculata and Casuarina equisetifolia, were screened for their biological properties.
3.1. Extraction yield
The yield of each extraction is mentioned in Table 1. Methanolic extraction of C. equisetifolia gave the highest yield
(8.99%), while the best extraction efficiency of O. corniculata was obtained with hydroethanolic extraction (13.18%).
The hydroethanolic and dichloromethane extraction of O. corniculata have a higher yield (13.18% and 4.66%)
compared to those of Casuarina equisetifolia (5.54% and 0.72%).
3.2. Antibacterial activity
Growth inhibitory effect of extracts: All extracts demonstrated antibacterial activity by inhibiting the growth of one
or more tested bacteria (Table 2). The hydroethanolic extract (10 mg/ml) of O. corniculata is the most active by
inhibiting all tested bacteria. The dichloromethane extract of O. corniculata also inhibits the growth of six bacteria
except S. abony. Similarly, the methanol and dichloromethane extracts of C. equisetifolia are also active by inhibiting
five bacteria out of seven. P. aeruginosa is the most sensitive bacteria its growth was inhibited by all the tested
extracts. S. abony is resistant to five extracts six. The active extracts in these trials were selected to determine their
minimum inhibitory concentrations (MIC).
Minimum Inhibitory Concentration (MIC): The Minimum Inhibitory Concentrations (MICs) of extracts are recorded in
Table 3. The extracts showed MIC values ranging from 0.078 to 5 mg/ml. Extracts obtained from O. corniculata
showed interesting antibacterial activity with MIC values ranging from 0.078 mg/ml to 5 mg/ml. The hydroethanolic
Table 1 yield for each extraction of C. equisetifolia and O. corniculata
C. equisetifolia O. corniculata
Solvants Plant material (g) % yield % yield
DM 100 0,72 4,66
Me 100 8,99 1,38
HE 50 5,54 13,18
DM: dichloromethane; Me: methanol; HE: Hydroalcoolic
Maceration:
It is a process of letting
stay solid material in a
liquid to extract
soluble compounds, or
solid material that
absorbs the liquid to
get the fragrance or
flavor.
CIP:
Collection Institute
Pasteur
Lagnika et al.
In vitro antibacterial activity of two medicinal plants used in Bénin to treat microbial infections,
Indian Journal of Science, 2014, 8(19), 10-15, www.discovery.org.in
http://www.discovery.org.in/ijs.htm © 2014 Discovery Publication. All Rights Reserved
Page
13
Table 2
Antibacterial activity of extracts at 10 mg/ml
Growth inhibition effect of extract at 10 mg/ml
Casuarina equisetifolia Oxalis corniculata
Extracts
Organisms DM ME H20/EtOH DM ME H20/EtOH
E. coli (CIP 53126) + + + - + -
S. aureus (ATCC 6538) - - + - - -
S. epidermidis - - + - - -
S.aureus methicillin resistant - - - - + -
E. faecalis (ATCC 2921) - - + - - -
P.aeruginosa (CIP 82118) - - - - - -
S. abony (CIP 8039) + + + + + -
DM: dichloromethane; ME: methanol; H
2
O/EtOH: Water/ethanol; -: inhibition; +: no inhibition
Table 3
Minimum Inhibitory Concentration of extracts from C. equisetifolia and O. corniculata extracts
Minimum Inhibitory Concentration (mg /ml)
Casuarina equisetifolia Oxalis corniculata
Extracts
Organismes DM MeOH H2O/EtOH DM MeOH H2O/EtOH
E. coli - - - 2.5 - 2.5
S.aureus > 5 2.5 - 2.5 2.5 5
S. epidermidis > 5 2.5 - 2.5 2.5 0.078
S.aureus MR 2.5 0.625 0.625 0.313 - 2.5
E.faecalis > 5 2.5 - 5 2.5 2.5
P. aeruginosa > 5 2.5 2.5 2.5 2.5 2.5
S. abony - - - - - 2.5
DM: dichloromethane; MeOH: methanol; H
2
O/EtOH: Water/ethanol; -: not active
Table 4
Total Activity of extracts
Total activity (ml/g)
Casuarina equisetifolia Oxalis corniculata
Extract from 1g (mg) 17.94 89.91 55.48 46.66 13.84 131.96
Extracts
Organismes DM MeOH H2O/EtOH DM MeOH H2O/EtOH
E. coli - - - 18.64 - 52.8
S. aureus <1.45 36 - 18.64 5.53 26.4
S. epidermidis <1.45 36 - 18.64 5.53 1689.7
S. aureus MR 3 144 89 149 - 52.8
E. faecalis <1.45 36 - 9.3 5.53 52.8
P. aeruginosa <1.45 36 22 18.64 5.53 52.8
S. abony - - - - - 52.8
DM: dichloromethane; ME: methanol; H
2
O/EtOH: Water/ethanol; -: not active
Lagnika et al.
In vitro antibacterial activity of two medicinal plants used in Bénin to treat microbial infections,
Indian Journal of Science, 2014, 8(19), 10-15, www.discovery.org.in
http://www.discovery.org.in/ijs.htm © 2014 Discovery Publication. All Rights Reserved
Page
14
and dichloromethane extracts of demonstrated highest inhibition
toward S. epidermidis and S.aureus Meticillin Resistant with MIC values
of 78 µg/ml and 313 µg/ml respectively. The MIC values obtained with
C. equisetifolia extracts range from 0.625 mg/ml to 5 mg/ml and the
best activities were obtained with methanol and hydroalcoolic extracts
with MIC value of 0.625 mg/ml against Staphylococus aureus meticillin
resitant (SARM). Overall, the antibacterial activity of extracts from O.
corniculata is higher than extracts from C. equisetifolia.
Total activity: The total activity of active extracts has been calculated
(Table 4). The extracts with higher total activity (TA) values are
considered the best. The most interesting total activities of O.
corniculata were obtained with dichloromethane (149 ml) and
hydroethanol (1689.7 ml) extracts against Staphylococus aureus
meticillin resistant (MRSA) and S. epidermidis respectively while the methanol (144 ml) and hydroethanol (89 ml)
extracts of C. equisetifolia showed best total activity against Staphylococus aureus meticillin resitant with (MRSA).
3.3. Artemia salina toxicity assay
Brine shrimp lethality test results are showed in Table 5. The LC50 values of the tested extracts ranged between 6.47
and 26.87 mg/ml. The highest LC50 were obtained with the dichloromethane extract of C. equisetifolia (LC50 = 22.02
mg/ml) and methanol extract of O. corniculata (LC50 = 26.87mg/ml). The hydroethanolic and methanol extracts of C.
equisetifolia and, hydroethanolic extract of O. corniculata were more toxic to shrimp with LC50 of 8.76, 6.47, and 8.16
mg/ml, respectively.
4. DISCUSSION
Medicinal plants are sources of antimicrobial agents, which can be exploited in the management of human diseases.
The plants are used medicinally in different countries of the world and are a good source of many potent and
powerful drugs (Mahesh and Satish, 2008). Resistance of pathogens to antibiotics and antifungal commonly used, the
increase in opportunistic infections and the effect of toxicity due to the continued use of several drugs have led to
increased attention paid to the search for new therapeutic agents from various sources, including plants, which are
good starting materials for the discovery of new antimicrobial agents (Sasidharan et al. 2011; Saad et al. 2011). In the
present study, biological activities of two medicinal plants of Benin pharmacopeia have been investigated.
The MIC values obtained with O. corniculata extracts range from 0.078 mg/ml to 5 mg/ml and the hydroethanolic
extract was the most active with MIC values of 0.078 mg/ml and 5 mg/ml against S. epidermidis and S. aureus
respectively. Similar results have been obtained with aqueous extract of O. corniculata (CMI = 6 mg/ml) against S.
aureus (Satish et al. 2008). MICs obtained with C. equisetifolia extracts range from 0.625 mg/ml to 5 mg/ml and
methanol and hydroethanolic extracts showed the best activities with MIC value of 0.625 mg/ml against
Staphylococus aureus meticillin resitant (SARM). Our results are more interesting than those obtained by Nehad and
Abdulrahaman (2012) in which the methanol extract showed a MIC of 50 mg/ml against the same bacteria. It is highly
probable that the antibacterial compounds of fruits of C. equisetifolia and leaves of O. corniculata are more soluble in
the hydroalcoholic solvent. This explains the strong antibacterial activity of hydroethanolic extract against Gram
positive and Gram negative bacterial strains. Usually, the use of alcohol as extractant is often encouraged to extract
antimicrobial compounds from plant materials (Selowa et al. 2010; Sati and Joshi, 2011; Olajuyigbe and Afolayan,
2012).
The methanol and hydroethanol extracts gave a MIC value of 62.5 μg/ml against Staphylococus aureus meticillin
resistant. This is in agreement with work by Ahsan et al. (2009), in which they obtained a MIC value of 64 µg/ml for
methanol extract of C. equisetifolia against S. aureus. These comparable results suggest that polar extracts of C.
equisetifolia are more active against of S. aureus strains. Brine shrimp lethality assay showed interesting activity with
LC50 values ranging from 6.47 to 26.87 mg/ml. These interesting results indicate that none of the extracts are toxic to
larvae. This is confirmed by the work of Zakaria et al. (2007) in which the author states that extracts are toxic when
the LC50 is less than 100 µg/ml. Thus, we concluded that all extracts tested in this study exhibited very low or no
toxicity, giving LC50 values higher than 100 μg/ml.
Table 5
Brine shrimp lethality assay of extracts from C. equisetifolia and
O. corniculata
Species Extracts LC
50
R
2
Casuarina
equisetifolia
DM 22.02 0.96
MeOH 8.76 0.94
H20/Et0H 6.47 0.90
Oxalis corniculata
DM 13.91 0.92
MeOH 26.87 0.98
H20/Et0H 8.16 0.95
DM: dichloromethane; MeOH: methanol; H2O/EtOH:
water/ethanol
Artemia salina:
It is a species of
crustacean lives in salt
lakes, lagoons and salt
marshes. It is used to
evaluate the
preliminary toxicity of
extracts, compounds
or wastewater.
SARM:
Staphyloccocus
aureus meticillin
resistant
Lagnika et al.
In vitro antibacterial activity of two medicinal plants used in Bénin to treat microbial infections,
Indian Journal of Science, 2014, 8(19), 10-15, www.discovery.org.in
http://www.discovery.org.in/ijs.htm © 2014 Discovery Publication. All Rights Reserved
Page
15
5. CONCLUSION
This study confirmed the traditional use of O. corniculata and C. equisetifolia and suggests that some of the extracts
having antibacterial properties can be further explored as a possible antibacterial agent source for the management
of infectious pathogenic diseases. The most important result was that hydroethanolic extract of O. corniculata
revealed significant antibacterial effect against S. epidermidis.
SUMMARY OF RESEARCH
This work was used to evaluate the antibacterial and preliminary toxicity of two medicinal plants. The findings give a scientific basis to the
traditional uses of O. corniculata and C. equisetifolia.
FUTURE ISSUES
It is still unknown which compounds are responsible for the biological activity of the two medicinal plants. Thus, bioassay-guided isolation and
identification of the active secondary metabolites of these plants will be process.
ACKNOWLEDGE
The authors are grateful to the medicinal plants seller and traditional practitioners from Ouémé and Ouidah regions. Helpful work of Botanist,
Dr. Yedomohan, from Herbier National of University of Abomey-calavi is appreciated.
DISCLOSURE STATEMENT
There is no financial support for this research work from the funding agency.
REFERENCE
1. Doughart JH, Okafor B. Anti-microbial activity of Senna
alata Linn. East and Central Afr J. Pharm. Sci., 2007, 10, 17-
21
2. George WB. A review of pathogenic microbiology Sai nt
Louis. CV Mosby Company. 1974
3. Soforowa A. Medicinal plants and traditional Medicine in
Africa. 1st ed. Spectrum Books Ltd., Ibadan, Nigeria, 1982
4. Ngo LT, Okogunb JL, Folk WR. 21st Century natural product
research and drug development and traditional medicines.
Nat. Prod. Rep., 2013, 30, 584-592
5. Kennedy DO, Wightman EL. Herbal extracts and
phytochemicals: Plant secondary metabolites and the
enhancement of human brain function. Adv. Nutr., 2011, 2,
32-50
6. Adedapo AA, Jimoh FO, Afolayan AJ, Masika PJ. Antioxidant
properties of the methanol extracts of the leaves and stems
of Celtis Africana. Records of Natural Products, 2009, 3, 23-
31
7. Bolou GEK, Attioua B, N’Guessan AC, Coulibaly A,
N’Guessan JD, Djaman AJ. Evaluation in vitro de l’activité
antibactérienne des extraits de Terminalia glaucescens
planch sur Salmonella typhi et Salmonella typhimurium.
Bulletin de la Société Royale des Sciences de Liège, 2011,
80, 772-790
8. Traoré Y, Ouattara K, Yéo D, Doumbia I, Coulibaly A.
Recherche des activités antifongique et antibactérienne des
feuilles d’Annona senegalensis Pers. (Annonaceae). Journal
of Applied Biosciences, 2012, 58, 4234- 4242
9. Eloff JN. A sensitive and quick method to determine the
minimal inhibitory concentration of plant extracts for
bacteria. Planta Med., 1998, 64, 711-713
10. Keymanesh K, Hamedi J, Moradi S, Mohammadipanah F,
Sardari S. Antibacterial, antifongique and toxicity of rare
Iranian plants. Int. J. Pharm., 2009, 5, 81-85
11. Eloff JN, Katerere DR, McGaw LJ. The biological activity and
chemistry of the southern African Combretaceae. J.
Ethnopharmacol., 2008, 119, 686-699
12. Eloff JN. Quantification the bioactivity of plant extracts
during screening and bioassay guided fractionation.
Phytomedicine, 2004, 11, 370-371
13. Mahesh B, Satish S. Antimicrobial activity of some
important medicinal plants against plant and human
pathogens. World J. Agric. Sci., 2008, 4, 839-843
14. Sasidharan S, Prema B, Yoga LL. Antimicrobial drug
resistance of Staphylococcus aureus in dairy products.
Asian Pac. J. Trop. Biomed., 2011, 1, 130-132
15. Saad S, Taher M, Susanti D, Qaralleh H, Rahim NABA.
Antimicrobial activity of mangrove pla nt (Lumnitzera
littorea). Asian Pacific journal of tropical medicine, 2011, 4,
523-525
16. Satish S, Raghavendra MP, Raveesha KA. Evaluation of the
Antibacterial Potential of some plants against human
pathogenic bacteria. Adv. Biol. Res., 2008, 2, 44-48
17. Nehad MG, Abdulrahaman SH. Antimicrobial efficacy of
Casuarina equisetifolia extracts against some pathogenic
microorganisms. J. Med. Plants Res., 2012, 6, 5819-5825
18. Selowa SC, Shai LJ, Masoko P, Mokgotho MP, Magano SR.
Antibacterial activity of extracts of three Croton species
collected in Mpumalanga region in South Africa. Afr. J.
Trad. CAM, 2010, 7: 98-103
19. Sati SC, Joshi S. Aspects of antifungal potential of
ethnobotanically known medicinal plants. Res. J. Med.
Plant, 2011, 5, 377-391
20. Olajuyigbe OO, Afolayan AJ. In vitro pharmacological
activity of the crude acetone extract of Erythrina caffra
Thunb: antibacterial and antifungal assessment. Journal of
Medicinal Plants Research, 2012, 6, 1713-1720
21. Ahsan R, Islam M, Haque E, Mossaddik A. In vitro
antibacterial screening and toxicity study of some different
medicinal plants. World Journal of Agriculture Sciences,
2009, 5, 617-621.
22. Zakaria HM, Mainen JM, Pax JM, Modest CK, Ramadhani
SON. Antimicrobial activity and brine shrimp toxicity of
extracts of Terminalia brownii roots and stem. BMC
Complementary and Alternatives Medicine, 2007, 7, 1-5
