EVALUATION OF THE ANTIFUNGAL ACTIVITY OF MORINGA OLEIFERA SEEDS, LEAVES AND FLOWERS

Article (PDF Available)inWorld Journal of Pharmaceutical Research 4(2):18 - 25 · January 2015with 3,793 Reads
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Abstract
Background: Moringa oleifera is one of multipurpose tree and rich in antimicrobial agents, that can be used for discovery of new classes of compounds that help in the treatment of microbial resistance. Materials and methods: seeds, leaves and flowers of Moringa were extracted by ether, ethanol, and water and tested for its antifungal activity using macrodilution method. Results: The nine extracts of Moringa oleifera were tested against 16 fungal isolates (Aspergillus flavus, A. fumigatus, A .terreus, A. niger, Candida albicans, C. glabrata, C. krusei, C. parapsilosis, one dermatophyte Trichophyton rubrum and Madurella mycetomatis). All of the tested organisms were resistant to seed oil type 1 and 2, leaves and flower of ether extract. It was also found to be resistant to leave water. The best extract was seed alcohol and seed water. All of the Moringa extracts irrespective of their types, in different concentrations did not inhibit the growth of all Candida species. KEYWORD: Moringa oleifera (seeds, leaves and flowers), phytoconstituents, minimum effective concentration (MEC), minimum inhibitory concentration (MIC). INTRODUCTION In Africa the use of traditional medicinal plants continue to form the basis of rural medicinal care due to the fact that these medicines are easy to obtain and cheap. The extensive use of antibiotics worldwide lead to increase in the microbial resistance which then lead to difficulty in the treatment of various diseases. In addition to the increased costs of hospital stay. Microbial resistance to drugs involved also fungal species. Previous studies reported the resistance of Candida species to azol drug to be as follows: Candida albicans 1.0%–2.1%, Candida parapsilosis 0.4%–4.2% and 1.4%–6.6% for Candida tropicalis.[1, 2] A plant known as Moringa oleifera was confirmed as multipurpose tree which have different uses in the treatment of various disease and as well possesses antibacterial and antifungal activity.[3] The aim of the present study was to assess the antifungal activity and to determine the minimum effective concentration of Moringa oleifera plant against 16 clinical fungal isolates.
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THE ANTIMICROBIAL ACTIVITY AND PHYTOCHEMICAL
CHARACTERISTIC OF MORINGA OLEIFERA SEEDS, LEAVES, AND
FLOWERS
Sahar M. Kheir1, Kafi S K1 and Haitham Elbir2*
1Department of Microbiology National Ribat University Hospital.
2Tropical Medicine Research Institute, Department of Microbiology. Khartoum-Sudan.
ABSTRACT
Background: Moringa oleifera was traditionally used in west Sudan
for purification of water. In this study, ether, alcohol and water extracts
of Moringa oleifera (seeds, leaves and flowers) were tested for their
antimicrobial effect against control strains and 155 clinical bacterial
isolates. The isolates included gram-negative bacteria (E. coli, P.
mirabilis, P. aeruginosa, K. Pneumoniae, Salmonella tyhimurium,
Enterobacter species, Salmonella typhi, Salmonella paratyphi B,
Shigella flexneri, Shigella sonnei and Serratia marcescens). While
gram-positive bacteria included (S. aureus, MRSA, S. epidermidis,
Diphtheroid, S. faecalis, S. pyogenes, L. monocytogenes and Streptomyces somaliensis).
Methods: The phytochemical screening of Moringa oleifera (seeds, leaves and flowers)
extracts was performed using qualitative determination whilst the antimicrobial activity of
ether, alcohol and water extracts of seeds, leaves and flowers was performed using agar
diffusion and macrobroth dilution method. Results: The results of the phytochemical analysis
demonstrated the presence of alkaloids, steroids, flavonoids, tannins and saponin. The studied
extracts displayed no activity on gram-negative bacteria whilst displayed various degrees of
antibacterial activities against gram-positive bacteria. The extract of seed alcohol, seed water
and leaf alcohol was active against all gram-positive. The extract of seed alcohol was very
active against S. aureus, Diphtheriod, with the lowest recorded minimal inhibitory
concentration (MIC) of 31.25mg/ml, while the seed water extract was very active against S.
aureus, Diphtheriod with lowest recorded MIC of 62. 5mg/ml. Conclusion: The results of the
present study support that Moringa oleifera has antibacterial activity against gram-positive
bacteria tested herein.
World Journal of Pharmaceutical Research
SJIF Impact Factor 5.045
Volume 4, Issue 01, 258-271. Research Article ISSN 2277 710541
Article Received on
03 Nov 2014,
Revised on 24 Nov 2014,
Accepted on 17 Dec 2014
*Correspondence for
Author
Sahar M. Kheir
Department of
Microbiology National
Ribat University Hospital.
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Elbir et al. World Journal of Pharmaceutical Research
KEYWORDS: antimicrobial, Moringa oleifera (leaves, seeds, flower), phytocomponents,
Minimum inhibitory concentration (MIC).
INTRODUCTION
Since ancient times and in early history, plants were and still important in treatment of
disease. The uses of plants continue to be witnessed throughout the world. The World Health
Organization (WHO) estimated that 80 % of the population of some developing countries
relies on herbal medicine for some aspect of primary health care.[1] Indigenous remedies are
the only form of therapy available to the majority of poor people. It has been estimated that
only 11% of the population has access to formal health care. [2]
Moringa oleifera is one of the 14 species of the family Moringaceae. [3] According to Muluvi
the Moringa tree was introduced to Africa from India at the turn of the twentieth century
where it was to be used as a health supplement. [4] M. oleifera is referred to as the ‘drumstick
tree’ or the ‘horse radish tree’, whereas in others it is known as the kelor tree. [5] While in the
Nile valley (Sudan), the name of the tree is ‘Shagara al Rauwaq’,which means ‘tree for
purifying. [6] M .oleifera has been widely used for the treatment of different types of diseases
due to its antibacterial activity. It is rich in compounds containing the simple sugar, rhamnose
and a unique group of compounds called glucosinolates and isothiocyanates. [7, 8] Other
medical proprieties include antipyretic, antiepileptic, antiinflamatory, antiulcerative,
antihypertensive, cholesterol lowering, antioxidant, antibacterial and antifungal activities, anti
diabetic, hepatoprotective. [9, 10, 11, 12, 13] M. oleifera had been tested before but the active
components of the plant may vary according to the geographical location. This study is
conducted to evaluate the antimicrobial effect, minimum inhibitory concentration and
phytochemical characteristic of seeds, leaves and flowers of M. oleifera.
MATERIALS AND METHODS
Preparation of the extract and phytochemical analysis of the plant
The plant used in this study (M. oleifera) was collected from a farm in Sennar State, Sudan
during the period from April to October 2013. This plant was taxonomically identified and
authenticated from the Medicinal and Aromatic Plants Research Institute (MAPPRI), in
Khartoum, Sudan. The flowers, leaves were dried in shade at room temperature and were
ground in a mortar to form powder. The brown shells of seeds were removed and the white
Kernels were ground in a mortar to form powder. Thereafter, 170g of each of the leaves,
seeds, and flower of Moringa were extracted by petroleum ether, ethanol and water
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successively by using soxhelet apparatous. The extract was filtered using Whatman filter
paper No (1) and evaporated under reduced pressure using Rota-vap. The extracted plant
material was then dried inside oven at 45°C to prevent contamination and to ensure that it is
completely evaporated. Each residue was weighed. The different extracts were poured in to
screw capped glass universal bottles and kept in refrigerator until used. The phytochemical
analysis was performed using qualitative determination according to. [14, 15, 16, 17]
Bacterial strains and Identification
The strains of E. coli ATCC25922, P. mirabilis ATCC35659, P. aeruginosa ATCC27853, K.
pneumoniae ATCC43816, Salmonella tyhimurium ATCC14028 and S. aureus ATCC25923
were provided by the bacteriology department of the national health laboratory. The bacterial
strains were isolated from 155 patients presenting with different clinical condition (wound
infection, urinary tract infection, peritonitis, tonsillitis, cough, enteric fever and exit site
infection) during 2013 -2014 in Khartoum state, Sudan. Specimens were aseptically collected
and inoculated on to chocolate agar, blood agar and MacConkey agar (Hi-Media, Mumbai
India). Once pure colonies identified, conventional tests were performed for identification
following standard procedures. [18, 19]
Antibiotic profile
Susceptibilities of control and clinical strain to, cefazolin, vancomycin, gentamicin,
ciprofloxcin, amikacin, ceftazidime, meropenem, ceftriaxone, penicillin and erythromycin
(Hi-Media, Mumbai India) were determined according to the Clinical and Laboratory
Standards Institute. [20]
Bacterial susceptibility to plant extract
The minimal inhibitory concentrations (MICs) of the three plant extracts were determined
using agar diffusion methods. Briefly, Bacterial culture adjusted to 0.5 McFarland standards
were inoculated into Muller & Hinton media (Hi-Media, Mumbai India) Four wells with
7mm in diameter were then cut in the inoculated culture media plate using sterile cork borer.
One gram of each extract was dissolved in 1ml of D.W for water extract and 1% dimethyl
sulfoxides (DMSO) for ether and alcohol extracts in separate tubes, then serially diluted two
fold to obtain final concentration (500mg/ml, 250 mg/ml, 125mg/ml, 62.5mg/ml, 31.25
mg/ml and 15.6 mg/ml). One hundred microliters of each prepared concentrations were then
added into the corresponding wells. The plates were then left at room temperature for 1 hour.
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Then incubated at 37°C for 24 h. Inhibition zone around each well were then measured using
a ruler in millimeters.
As for the oil extract, Four concentrations (100%, 75%, 50%, 25%) v/v of Moringa seed oil
type 1 (oil extracted by ether), type 2 (ready to use oil) was prepared as follows: one colony
of bacteria was emulsified in 1ml of undiluted seed oil (100%), for the remaining
concentrations the 75 µl, 50 µl and 25µl of oil was mixed with 25 µl, 50 µl, 75 µl of bacterial
broth culture adjusted to 0.5 McFarland standards respectively. Five drops of ether was added
to each tube to help in emulsification. The mixture was incubated at 35ºC for 24h. Each of the
tests mentioned above was performed in duplicates. The MIC of the oil was determined by
sub-culturing samples from the concentrations above on new plates of Mueller Hinton agar at
35ºC for 48h. [20, 21]
Susceptibility testing of the aerobic actinomycetes (Streptomyces species.) was done using
broth macrodilution method. The colonies were dissolved in Brain heart infusion broth and
vortexed for 15 seconds. Then adjusted to 0.5 Macfarland standards to obtain approximately
5×105 CFU/ml. This dilution was achieved when 500µl from the adjusted inoculums was
added to 500µl of the prepared extracts. Serial dilution was then done to achieve the
following concentration (250 mg/ml, 125mg/ml, 62.5mg/ml, 31.25 mg/ml 15.6 mg/ml, and
7.8 mg/ml) in a sterile test tube. The seed oil was done as method mentioned above. Then
was incubated at 37 ± 2 ºC for three days, The MIC was taken as the lowest concentration
that prevented bacterial growth. To exclude contamination of plant extract, the essay was
performed as above without inoculation of organism. To exclude the solvent effect the essay
was performed as above with the solvent and bacterial strain without adding the plant
extract.[22]
RESULTS
Three parts of Moringa oleifera were phytochemically analysed and they showed variation in
their chemical component as shown in table 1
Table 1: Phytochemical component of the different parts of M. oleifera extracts
Seeds
Flavonoids
Alkaloids
Steroid
Tanin
Glycosides
Reduced sugars
Ether extract
N
P
P
N
N
N
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Alcoholic extract
P
P
N
P
P
N
Aqueous extract
P
N
N
T
N
N
Leaves
Ether extract
N
P
P
N
N
N
Alcoholic extract
P
N
T
P
N
N
Aqueous extract
T
N
N
T
N
N
Flowers
Ether extract
N
T
P
N
N
N
Alcoholic extract
T
T
P
T
N
N
Aqueous extract
T
N
N
T
N
N
*P: Positive, N: Negative, T: Trace.
All extracts from the different parts of Moringa showed no anti-microbial activity against the
tested gram negative bacteria. The seed alcohol extracts exerted antimicrobial activity against
all the tested gram positive bacteria with variation of its action according to concentrations. It
showed inhibition zones for all gram positive at concentration of 62.5 mg/ml except 3
organisms Diphtheroid, S. aureus against which it was effective up to concentration of
31.23mg/ml and S. pyogenes, L. monocytogenes up to 125mg/ml. The best action of the
extract was against the standard S. aureus inhibition zone of (28mm at concentration of
500mg/ml) table 2A and 3. Seed water was the second effective extract against gram positive
organisms table 2B. In this study M. oleifera seed oil type1, type 2 extracted were used at
concentrations 100%, 75%, 50%, 25% revealed no antibacterial activity against gram positive
bacteria control strains and clinical isolates and so leaves and flowers ether at (500mg/ml).
Table 2A: The inhibition zones of the different concentrations of alcohol extract of M.
oleifera seed on bacterial isolates. Diameter of the inhibition zone in mm
Bacteria species
No
500
mg/ml
250
mg/ml
125 mg/ml
62.5 mg/ml
31.25
mg/ml
15.6
mg/ml
Staphylococcus aureus
ATCC
1
28
22
10
8
6
resistant
Staphylococcus aureus
36
27-24
22-18
16-12
10-7
6-3
resistant
Methicillin-resistant
Staphylococcus aureus
(MRSA)
3
24-22
20-17
15-13
9-7
resistant
resistant
Staph. epidermidis
12
25-22
18-16
15-12
11-3
resistant
resistant
Diphtheroid speices
12
27-24
22-19
17-13
11-8
6-3
resistant
Streptococcus faecalis
5
22-20
20-17
15-12
11-9
resistant
resistant
Streptococcus pyogenes
2
24-22
20- 16
12-10
resistant
resistant
resistant
Listeria monocytogenes
1
13
11
8
resistant
resistant
resistant
Table 2B: The inhibition zones of the different concentrations of water extract of M.
oleifera seed on bacterial isolates. Diameter of the inhibition zone in mm
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Table 2D showed that the leaves water, showed no activity against S. pyogenes, L.
momocytogenes, and Diphtheriod species.
Two isolates of S. aureus (isolated from urine) were resistant to all Moringa parts. While 3
isolates of S. epidermidis isolated from peritoneal fluid and exit site infection of peritoneal
catheter, and 5 isolates of Diphtheroid species isolated from exit site were resistant to leaves
alcohol at 500mg/ml. Leaves ether showed inhibitory action only against Diphtheroid, S.
pyogenes, L. momocytogenes. The Diphtheroid was sensitive to leaf ether extract at
500mg/ml with zones of inhibition (17-16mm), 250mg/ml (14-10mm), 125mg/ml (10-7mm)
and 62.5mg/ml (6-2mm). The MIC of leaf ether to Diphtheroid was 62.5mg/ml. S. pyogenes
was inhibited at the same concentrations above with inhibition zones of 16-14mm, 12-9 mm,
8-7 mm and 6-3mm respectively and MIC of 62.5mg/ml. For L. momocytogenes the
inhibition zone were 20mm, 14mm and 8mm for the concentration 500mg/ml, 250mg/ml and
125mg/ml respectively. The MIC was 125mg/ml.
Table 2C: The inhibition zones of the different concentrations of alcohol extract of M.
oleifera leaf on bacterial isolates. Diameter of the inhibition zone in mm
Bacteria species
500 mg/ml
250 mg/ml
125
mg/ml
62.5 mg/ml
31.25
mg/ml
15.6
mg/ml
Staphylococcus aureus
ATCC
25
16
7
resistant
resistant
resistant
Staphylococcus aureus
25-20
18-15
12-3
resistant
resistant
resistant
Methicillin-resistant
Staphylococcus
aureus(MRSA)
20-19
16-10
8-5
resistant
resistant
resistant
Staph. epidermidis
22-18
15-10
8-4
resistant
resistant
resistant
Diphtheroid speices
27-23
20-18
15-10
8-3
resistant
resistant
Streptococcus faecalis
18-15
13-10
9-3
resistant
resistant
resistant
Bacteria species
500
mg/ml
250 mg/ml
125 mg/ml
62.5
mg/ml
31.25mg/ml
15.6 mg/ml
Staphylococcus aureus ATCC
24
19
12
8
resistant
resistant
Staphylococcus aureus
24-21
19-17
14-11
10-4
resistant
resistant
Methicillin-resistant
Staphylococcus aureus
(MRSA)
20-17
16-14
7-0
resistant
resistant
resistant
Staph. epidermidis
22-18
14-10
9-6
resistant
resistant
resistant
Diphtheroid speices
22-17
15-12
10-7
6-2
resistant
resistant
Streptococcus faecalis
25-22
20-17
14-4
resistant
resistant
resistant
Streptococcus pyogenes
23-20
17-14
13-8
resistant
resistant
resistant
Listeria monocytogenes
15
14
10
resistant
resistant
resistant
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Streptococcus pyogenes
24-22
21-13
12-7
resistant
resistant
resistant
Listeria monocytogenes
12
10
resistant
resistant
resistant
resistant
Table 2D: The inhibition zones of the different concentrations of water extract of M.
oleifera leaf on bacterial isolates. Diameter of the inhibition zone in mm
Table 2E, 2F showed that the flower alcohol and flower water extracts showed no activity
against S. pyogenes and L. momocytogenes.
Table 2E: The inhibition zones of the different concentrations of alcohol extract of M.
oleifera flower on bacterial isolates. Diameter of the inhibition zone in mm
Bacteria species
500
mg/ml
250 mg/ml
125 mg/ml
62.5 mg/ml
31.25
mg/ml
15.6
mg/ml
Staphylococcus aureus
ATCC
23
16
10
resistant
resistant
resistant
Staphylococcus aureus
23-21
18-16
12-6
resistant
resistant
resistant
Methicillin-resistant
Staphylococcus
aureus(MRSA)
22-20
18-15
11-4
resistant
resistant
resistant
Staph. epidermidis
20-15
12-10
6-4
resistant
resistant
resistant
Diphtheroid speices
20-18
16-13
9-5
resistant
resistant
resistant
Streptococcus faecalis
22-16
15-12
10-5
resistant
resistant
resistant
Streptococcus pyogenes
resistant
resistant
resistant
resistant
resistant
resistant
Listeria monocytogenes
resistant
resistant
resistant
resistant
resistant
resistant
Table 2F: The inhibition zones of the different concentrations of water extract of M.
oleifera flower on bacterial isolates. Diameter of the inhibition zone in mm
Bacteria species
500
250 mg/ml
125 mg/ml
62.5 mg/ml
31.25
15.6 mg/ml
Bacteria species
500
mg/ml
250
mg/ml
125
mg/ml
62.5 mg/ml
31.25
mg/ml
15.6 mg/ml
Staphylococcus aureus
ATCC
20
15
10
resistant
resistant
resistant
Staphylococcus aureus
20-17
14-11
10-3
resistant
resistant
resistant
Methicillin-resistant
Staphylococcus
aureus(MRSA)
19-17
13-10
resistant
resistant
resistant
resistant
Staph. epidermidis
20-18
14-7
6-4
resistant
resistant
resistant
Diphtheroid speices
resistant
resistant
resistant
resistant
resistant
resistant
Streptococcus faecalis
17-12
10-7
6-4
resistant
resistant
resistant
Streptococcus pyogenes
resistant
resistant
resistant
resistant
resistant
resistant
Listeria monocytogenes
resistant
resistant
resistant
resistant
resistant
resistant
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mg/ml
mg/ml
Staphylococcus aureus
ATCC
20
15
10
resistant
resistant
resistant
Staphylococcus aureus
20-15
12-9
8-3
resistant
resistant
resistant
Methicillin-resistant
Staphylococcus aureus
(MRSA)
17-15
12-9
7-4
resistant
resistant
resistant
Staph. epidermidis
21-14
11-6
9-3
resistant
resistant
resistant
Diphtheroid speices
18-14
10-8
7-3
resistant
resistant
resistant
Streptococcus faecalis
15-13
11-6
5-3
resistant
resistant
resistant
Streptococcus pyogenes
resistant
resistant
resistant
resistant
resistant
resistant
Listeria monocytogenes
resistant
resistant
resistant
resistant
resistant
resistant
The Minimum inhibitory concentration of each organism against the different extracts range
from 250mg/ml 31.25 mg/ml table 3.
Table 3: Minimum inhibitory concentration of Petroleum ether, Ethanol and Water
extract of M. oleifera seeds, leaves and flowers activity against human pathogens
expressed in mg/ml.
SA: seed alcohol, SW: seed water, LE: leaf ether, LA: leaf alcohol, LW: leaf water, FA:
flower alcohol, FW: flower water.
Streptomyces was completely inhibited by seed water and resistant to seed oil type 1 and 2
leaves ether, leaves water, flower ether as shown in (table 4).
Table 4: Antimicrobial activity of Petroleum ether, Ethanol and Water extract of M.
oleifera seeds, leaves and flowers activity against Streptomyces somaliensis.
Clinical isolates
SA
SW
LE
LA
L W
FA
F W
Staphylococcus aureus ATCC
31.25
62.5
resistant
125
125
125
125
Staphylococcus aureus
31.25
62.5
resistant
125
125
125
125
Methicillin-resistant
Staphylococcus aureus (MRSA)
62.5
125
resistant
125
250
125
125
Staph. epidermidis
62.5
125
resistant
125
125
125
125
Diphtheroid speices
31.25
62.5
62.5
62.5
resistant
125
125
Streptococcus faecalis
62.5
125
resistant
125
125
125
125
Streptococcus pyogenes
125
125
62.5
125
resistant
resistant
resistant
Listeria monocytogenes
125
125
125
250
resistant
resistant
resistant
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Extract type
Concentration required in mg/ml
250
125
62.5
31.25
15.6
7.8
Seed alcohol
No growth
No growth
No growth
No growth
Hazy
Moderate
Seed water
No growth
No growth
No growth
No growth
No growth
No growth
Leaf ether
Resistant
Resistant
Resistant
Resistant
Resistant
Resistant
Leaf alcohol
No growth
No growth
No growth
Hazy
Moderate
Heavy
Leaf water
Resistant
Resistant
Resistant
Resistant
Resistant
Resistant
Flower ether
Resistant
Resistant
Resistant
Resistant
Resistant
Resistant
Flower alcohol
No growth
No growth
No growth
Hazy
Moderate
Heavy
Flower water
No growth
No growth
No growth
No growth
Hazy
Clumpy
The appropriate standard antibiotic discs were tested against standard bacteria and clinical
isolates. All the organisms tested were sensitive to antibiotics used except MRSA was
resistant to all antibiotic tested except vancomycin and presented in Table 5A and 5B.
Table 5A: Antimicrobial activities of control strains and clinical isolates of gram
positive bacteria against the corresponding standard antibiotics.
Bacteria species
VA
CZ
G
CIP
CAZ
P
E
CTR
S. aureus ATCC
26
30
25
28
ND
ND
ND
ND
S. aureus
18-25
20-24
22-30
22-26
ND
ND
ND
ND
Methicillin-resistant
Staphylococcus aureus (MRSA)
18-20
R
R
R
R
ND
ND
R
S. epidermidis
20-30
20-25
22-29
22-24
ND
ND
ND
ND
Diphtheriod species
25-31
ND
26-30
22-28
ND
ND
ND
ND
S. faecalis
20-24
R
18-20
22-25
ND
ND
ND
ND
S. pyogenes
ND
ND
ND
ND
ND
29-32
22-23
28-30
L. monocytogenes
ND
ND
30
32
ND
30
ND
R
VA: vancomycin, CZ: cefazolin, G: gentamicin, CIP: ciprofloxacin, CAZ: ceftazidime,
P: penicillin, E: erythromycin, CTR: ceftriaxone ND: not done, R: resistant, numbers
indicate the measurement of inhibition zone in mm
Table 5B: Antimicrobial activities of control strains and clinical isolates of gram
negative bacteria against the corresponding standard antibiotics.
Bacteria species
N
G
CIP
AK
CAZ
MRP
Escherichia coli ATCC
1
22
30
20
32
30
Klebsiella pneumoniae ATCC
1
22
38
25
32
30
Pseudomonas aeruginosa ATCC
1
24
29
23
26
29
Salmonella tyhimurium ATCC
1
ND
30
ND
ND
ND
Proteus mirabilis ATCC
1
23
25
30
33
30
P. aeruginosa
21
22-30
26-31
25-30
20-27
26-30
E. coli
27
20-30
28-30
20-26
26-30
25-35
Proteus mirabilis
13
25-30
22-30
18-20
22-25
25-32
K. Pneumoniae
8
15-22
21-28
18-22
30-35
22-32
Enterobacter species
2
32
23
25
22
25
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Salmonella typhi
1
ND
30
ND
ND
ND
Salmonella paratyhi B
1
ND
26
ND
ND
ND
Shigella sonnei
1
ND
22
ND
ND
ND
Shigella flexneri
1
ND
23
ND
ND
ND
Serratia marcescens
1
25
30
25
30
ND
G: gentamicin, CIP: ciprofloxacin, AK: amikacin, CAZz: ceftazidime, MRP:
meropenem, ND: not done, numbers indicate the measurement of inhibition zone in
mm.
DISCUSSION
Each of the extract tested in the current study showed no antibacterial activity on gram-
negative bacterial strains tested. However they showed antibacterial activities against gram-
positive bacterial strains. In general the seed alcohol, water extract showed the best
antibacterial activity followed by leaves alcohol extract against all grams positive bacteria
tested. The results of this present research showed that Moringa seed ethanol extract had
broadest spectrum of activity on the tested bacteria. The overall data of this study were in
accordance with previous results.
The antibacterial activity of Moringa seeds had been reported by many researchers. Our
results were in agreement with Saadabi who reported that Moringa seeds extract has the best
antibacterial activity. The petroleum ether had no antibacterial activity against S. aureus, E.
coli, P. aeruginosa while seed water had antibacterial activity against S. aureus. [23]
Some food pathogen like S. aureus, E. coli, were tested against seed ethanol extract and
found to be sensitive at 50mg/ml while Enterobacter spp, Shigella spp, P. aeruginosa, and
Salmonella typhi were not sensitive to seed ethanol extract. [24]
Also our findings are consistent with the findings of Gomashe who reported that the
Petroleum extract of leaves was not effective against all the test pathogens (Salmonella typhi,
Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus) at 30mg/ml except P.
aeruginosa (12mm). [25] Doughari 2007 who reported in their study on the antibacterial
activity from the aqueous, acetone and ethanol extracts of the leaves of M. oleifera that
ethanol extract of the plant demonstrated the highest activity, while the aqueous extract
showed the least activity at 100 mg/ml. [26] In this study leaf aqueous extract showed the least
activity and did not inhibit L. monocytogenes, S. pyogenes and Diphtheriod species, while
the ethanol extract showed significant and better activity at 125mg/ml.
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To the best of our knowledge, the antimicrobial activity of seed water against streptomyces
somaliensis described here was not recorded before. The overall antibiotic susceptibility
results were similar to antimicrobial activity of the plant extracts.
Ellert and Guevera reported that the antimicrobial activity of Moringa seed is due to the
presence of phyto-chemical compounds. Due to presence of 4(α-L-rhamnosyloxy)-benzyl
isothiocyanate. [27, 28] The concentrations of these phytochemicals and the variations in the
findings of this study compared to earlier studies reported may be due to the environment,
geographical site, genetics, and soil. [29] Moringa leaves ethanol revealed presence of
flavonoids and saponins and Moringa seed ethanol contains alkaloids in agreement with
bukar. [24] Moringa seed ethanol contain tannins and saponins in agreement with Napolean.
[30] The flowers contain pterogospermin, this component an active antibacterial activity. [31]
It has been reported that different solvents have different extraction capabilities. [25, 32, 33]
Himal reported that to extract broad spectrum antimicrobial compound from plant is
confirmed by the ethanol solvents. [34] The differences observed between antibacterial
activities of the extracts could be explained by the differences in the chemical composition of
these extracts.
CONCLUSION
M. oleifera extract proved to be highly potent against different strains of gram positive
organisms. Leaves and flowers possess antibacterial to a lesser extent. The best activity
reported by the seeds alcohol and water extract against S. aureus control strain and clinical
isolates. The inhibition zones of the plant are closely resembled to the inhibitory zones of the
antibiotics so can lead to the development of new chemical compounds which can be used to
treat various types of infection.
ACKNOWLEDGEMENT
The author would like to thank the staff member if the National Ribat University Hospital
and Peritoneal Dialysis Program Khartoum, Sudan for their great help.
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    The qualitative and quantitative phytochemical screening and antifungal activity of root and seed extracts of Moringa oleifera was investigated. Ethanolic, methanolic, hot water and cold-water extracts of Moringa oleifera were tested against Candida albicans. Results showed the presence of carbohydrate, tannins, saponins, cardiac glycosides, alkaloids and phytic acid while flavonoids, anthraquinone and steroids appeared negative. Also, quantitative estimations of 0.57% and 1.17% oxalates, 4.02% and 2.66% phytic acid, 1.92% and 3.99% alkaloid, 65.47mg/100g and 109.12mg/100g tannins for the roots and seeds respectively was recorded. Activity of the methanolic extracts was highest while the cold-water extracts had the least activity of all tested extracts. Moringa oleifera is a promising plant for use in antifungal infections of Candida albicans.
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