ISSN 2088-6586

Nwaiwu N.E., Ibrahim W.I. and Raufu I.A., 2012. Antiseptic and Coagulation Properties of Crude Extracts of
Moringa Oleifera Seeds from North East of Nigeria.
V o l.u m e 1 , N u m b e r 2 : 5 1 - 5 9 , F e b r u a r y , 2 0 1 2
T2012 Department of Environmental Engineering
Open Access http://www.trisanita.org/japes

Research Paper

ANTISEPTIC AND COAGULATION PROPERTIES OF CRUDE EXTRACTS OF

MORINGA OLEIFERA SEEDS FROM NORTH EAST OF NIGERIA
NWAIWU N.E.1*, IBRAHIM W.I.2 and RAUFU I.A.3
1Department

of Civil and Water Resources Engineering, 3Department of Veterinary Microbiology, Faculty

of Veterinary Medicine, University of Maiduguri, Maiduguri, Borno State, Nigeria.

2Department

of Hospital Management, Plateau State Polytechnic, Barkin Ladi, P.M.B. 2023,

Plateau State, Nigeria.

*Corresponding Author: Phone: +2348025528200; E-mail: nknwaiwu@yahoo.co.uk

Received: 21st June 2011; Revised: 12th July 2011; Accepted: 2nd January 2012

Abstract: The aim of this paper is to determine the difference between the antimicrobial
and coagulation activity of crude extracts of Moringa oleifera grown in different parts of
North East, Nigeria. Moringa oleifera seeds were plucked from trees in three states of
the North East Nigeria namely Adamawa, Borno and Yobe States. The results indicated
that crude aqueous extracts of Borno Moringa oleifera, Yobe Moringa oleifera and
Adamawa Moringa oleifera showed different degrees of growth inhibitions to
microorganisms. Statistically the origin of Moringa oleifera seed had no significant
influence on its antimicrobial properties from the present work. On coagulation the
Moringa oleifera seed from Adamawa State had the fastest turbidity removal rate of
18NTU/minute within the first 30minutes. This was faster than 12.4NTU/minute and
15.53NTU/minute for Moringa oleifera samples from Yobe and Borno states
respectively within the same time interval.
Keywords: Antimicrobial and coagulation activity, aqueous extracts, growth inhibitions

INTRODUCTION
Many plants products such as crude extracts have been explored for their antimicrobial
properties [1-9].Plant are rich sources of natural substances that can be utilized in the
development of environmentally and humanly safe methods for water treatment. Crude plants
extracts often consist of complex s of active compounds and they may show greater overall
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Nwaiwu N.E., Ibrahim W.I. and Raufu I.A., 2012. Antiseptic and Coagulation Properties of Crude Extracts of
Moringa Oleifera Seeds from North East of Nigeria.
.

bioactivity when compared to the individual constituents [10-12]. The screening of specific parts
of a plant such as the seeds, barks, roots, leaves and other parts for their extracts have yielded
excellent results on the bioactive constituent of such [13-15].
The Moringa oleifera seed which is noted as a good coagulant and softener has been found
to have antimicrobial properties [16-21]. Aqueous extracts of Moringa oleifera leaves posses
antimicrobial activity against E. coli, Klebsiella aerogenes, Klebsiella pneumoniae, S. aureus and
B. subtilis [22]. An active antimicrobial ingredient 4 & L rhamnosyloxy- benzyl isothiocyanate
has been identified by [23] in Moringa oleifera seeds. Moringa oleifera seeds treat water on two
levels acting both as a coagulant and an antimicrobial agent [24]. Crushed seeds of Moringa
oleifera clarify and purify river water to suit domestic uses and lower the bacterial concentration of
the water making it safe for drinking [25]. The aim of this paper is to determine the difference (if
any) between the antimicrobial and coagulation activity of crude extracts of Moringa oleifera
grown in different parts of North East, Nigeria.
MATERIALS AND METHODS
Study Area
Moringa oleifera seeds were plucked from trees in three states of the North East Nigeria
namely Adamawa, Borno and Yobe States. Adamawa State is located in the Northern Guinea
Savannah while Borno and Yobe reside in the Sudan Savannah.
Hexane Moringa oleifera oil and crude aqueous extracts
Moringa oleifera essential oils was extracted using hexane from twenty five (25grams), twelve
(12grams) and twenty (20grams) of Moringa oleifera crushed seed kernel respectively from
Borno, Yobe and Adamawa states. The debris resulting from the hexane oil extraction were
employed for the water extraction. Seven hundred milliliters (700ml) of water was added to each
debris sample and filtered. The filtrates were put on evaporating disks, inserted into a hot oven at
40oC 50oC for up to 24hours. The percentage yields of crude extracts from the seeds were as
follows: Borno (21.56%), Yobe (51%) and Adamawa (21.85%).
Water Extraction
The debris resulting from the hexane extraction of Moringa oleifera seed powder were used
for water extraction. Seven hundred millilitres (700ml) of water was added to each sample. The
contents of the thimbles were mixed and filtered. The filtrates were put on evaporating disks
inserted into a hot air oven at 40C -50C for up to 24hours.
Preparation of Media and Isolation of Microrganisms: Nutrient broth
After the identification of the organisms from the turbid water in the media they were mass
produced in the nutrient broth before the test of the extracts on the microorganism would be
carried out. Nutrient broth was weighed (9.1grams) and diluted in 700ml of distilled water. The
mixture was heated using a spirit lamp to dissolve the media. McConkey broth (single and double
strength) was prepared and dispensed into bottles with proper labels. The bottles and contents
were sterilized in the autoclave at 121C for 15mins. The water sample was innoculated into the
bottles accordingly and incubated at 35-37C for a minimum duration of 14hours. The bottles that
had positive microbial growth turned from purple to light purple (in the case of medium growth) or
to pale yellow (in the case of extreme growth).
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Nwaiwu N.E., Ibrahim W.I. and Raufu I.A., 2012. Antiseptic and Coagulation Properties of Crude Extracts of
Moringa Oleifera Seeds from North East of Nigeria.
.

Subculture Media
The Desoxycholate Citrate Agar (DCA), Eosin Methylene Blue Agar (EMBA and Salmonella
Shigella Agar (SS-Agar or SSA) media were prepared for subculturing and proper identification of
the organisms from the McConkey broth test. The Agar are poured into petri dishes and allowed
to cool. Using a sterilized wire loop, samples were taken from the positive tubes and streaked
across all the prepared media. The wire loop was sterilized before streaking each media. After
streaking, the petridishes were put into the incubator at 35C 37C for a minimum of 16hours.
After 24hours, Enterobacter was identified on the SS Agar with a pinkish colony while Salmonella
and Shigella were present as colourless. In DCA Salmonella and Shigella were also identified
with a colourless colony while in EMBA the duo formed a black and white colony.
Administration of Extracts on Organisms
The agar diffusion method was adopted for this study .Two grams (2g) of the water extract
was mixed with 10ml of distilled water. This was to allow the solid extract to dissolve to
suspension form. Each of the identified microorganisms (nutrient broth cultures) were introduced
into petri dishes containing already prepared Nutrient agar. The broth culture was poured over the
surface of the nutrient agar ensuring a proper spread over the entire surface. The petri dishes
and their content were left to stand for sometime after which the excess liquid was drained off.
Holes were made in each plate (8mm in diameter) using agar borer. Equal volumes of the
extracts were transferred into the holes in each petri dish. The plates were incubated for 24hours.
At the end of the incubations period, the plates were collected and zones of inhibition that
developed were measured.
Extracts Preparation for Coagulation
Turbid water samples were collected from a pond on the outskirts of Maiduguri. One liter of
the water was dispensed into 3 transparent plastic containers. A measure of 1.8g each of the
Moringa oleifera seed powder from Borno, Yobe and Adamawa States were mixed separately in
75ml of water. The aqueous mixture were shaken vigorously for 10minutes and gently for 5
minutes. The aim of the agitation was to release the polyelectrolytes responsible for coagulation.
The filtrates were obtained and dispensed into the samples of turbid water stirring all the time.
Supernatants were collected after 30mins, 1hr, 3hr, 6 hours and 24hours and tested for Total
dissolved solids (TDS), temperature , electrical conductivity, colour, pH and turbidity.
RESULTS AND DISCUSSION
Comparison of antimicrobial properties
The results indicated that crude aqueous extracts of Borno Moringa oleifera, Yobe Moringa
oleifera and Adamawa Moringa oleifera showed different degrees of growth inhibition (see Table
1). The aqueous extract of Borno Moringa oleifera showed the broadest antibacterial activity
against Salmonella (the diameter of inhibition zone, 15mm). The extracts from Yobe Moringa
oleifera and Adamawa, Moringa oleifera inhibited growth of Salmonella to the same degree (the
diameter of inhibition, 11mm). Equal zones of inhibition (12mm) against Shigella resulted from
Borno and Adamawa Moringa oleifera. The three types of Moringa oleifera seeds produced
inhibitory actions against Enterobacter, 11mm for Borno, 11mm for Yobe and 12mm for
Adamawa states. The antimicrobial properties of the Moringa oleifera seed is a desirable tool in
the control of undesirable microorganism in water treatment.
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Journal of Applied Phytotechnology in Environmental Sanitation, 1 (2): 51-59.

Nwaiwu N.E., Ibrahim W.I. and Raufu I.A., 2012. Antiseptic and Coagulation Properties of Crude Extracts of
Moringa Oleifera Seeds from North East of Nigeria.
.

Table 1: Zones of inhibition for Moringa oleifera seed powder crude extracts from Borno, Yobe
and Adamawa States.
Zones of inhibition (mm)
Origin of seed
Salmonella
Shigella
Enterobacter
Borno State

15

12

11

Yobe State

11

11

11

Adamawa State

11

12

15

A 2-way analysis of variance (ANOVA) without replication shows that the Moringa oleifera
seeds have no statistically significant difference in the individual microbial zones of inhibition.
This means that the Moringa oleifera seed powder exhibited equal inhibitory action against
Salmonella, Shigella and Enterobacter. (F=1.09, Fcrit=6.944, p=0.42). Similarly, the analysis also
did not show any significant difference in the actions against the microorganism in relation to the
place of origin (F= 0.4, Fcrit =6.944; p=0.69): This indicates that the origin of Moringa oleifera seed
has no influence on its antimicrobial properties from the present work.

total dissolved solids

values (mgL-1)

Comparison of Coagulation Properties

The values for the three Moringa oleifera samples are shown in Figures 1-3. The pH values
for the water sample using Moringa oleifera seed from Yobe State varied between 7.50 8.13.
The respective pH values for water treated with Moringa oleifera from Adamawa and Borno state
ranged between 7.50 -8.19 and 7.50-8.10. It can be observed that the pH of the treated water
remained within the allowable limits of the World Health Organization[26,27].This is in agreement
with [17] that noted that on softening water with Moringa oleifera seed powder, the pH was within
the WHO recommendation standards. Similar reports have also been made by [28] and [29].The
electrical conductivity values showed slight fluctuations within the twenty four hour period of the
experiment. After 24 hours, the Yobe Moringa oleifera sample water had a final value of 540
s/cm, from an initial value of 511s/cm. The final 24 hour values for Adamawa and Borno are
553 and 536 s/cm respectively.
An increment in the value of the total dissolved solids was observed from the treatment
employing all the Moringa oleifera seed samples. This increment may be due to the presence of
Moringa oleifera solid itself in the water(see Figs.1-3).
280
275
270
265
adamawa
260
borno
255
yobe
250
0
200
400
600
800
1000
1200
1400
1600
Contact time(mins)
Fig. 1: Effect of contact time on total dissolved solids
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Journal of Applied Phytotechnology in Environmental Sanitation, 1 (2): 51-59.

Nwaiwu N.E., Ibrahim W.I. and Raufu I.A., 2012. Antiseptic and Coagulation Properties of Crude Extracts of
Moringa Oleifera Seeds from North East of Nigeria.
.

8.3
8.2
8.1
pH

8
7.9
adamawa
borno
yobe

7.8
7.7
7.6
7.5
0

200

400

600

800

1000

1200
1400
1600
Contact time(mins)

Electrical conductivity S/cm

Fig. 2: Effect of contact time on pH

555
550
545
540
535
530
525
520
515
510

adamawa
borno
yobe
0

200

400

600

800

1000

1200

1400

1600

Contact time(mins)

turbidity removal efficiency(%)

Fig. 3: Effect of contact time on electrical conductivity

105
100
95
90
85
80
75
70
65
60

percentage rem(adamawa)
percentage rem(borno)
percentage rem(yobe)
0

200

400

600

800

1000

1200

1400

Contact time(mins)
Fig. 4: Effect of contact time on turbidity removal
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Journal of Applied Phytotechnology in Environmental Sanitation, 1 (2): 51-59.

1600

Nwaiwu N.E., Ibrahim W.I. and Raufu I.A., 2012. Antiseptic and Coagulation Properties of Crude Extracts of
Moringa Oleifera Seeds from North East of Nigeria.
.

Turbidity removal is a very important aspect of water treatment. Varying turbidity removal
efficiencies have been displayed by the Moringa oleifera samples used. Within an interval of
30mins the Moringa oleifera from Yobe state reduced the turbidity by 64.9% (see Fig, 4). This
trend continued through the 24hour period. At the end of 24hour, the turbidity removal efficiency
obtained was 91.34%. The Moringa oleifera powder from Adamawa State yielded a turbidity
reduction of 96.57% within a 30minute interval. This efficiency increased to 97.05% in one hour.
After 6hours, the removal efficiency was 98.2%. However, reduction of the efficiency was
experienced after a 24hour period (97.66%). The Moringa oleifera sample from Borno state had
30min turbidity reduction of 81.33% as well as a 24hour removal efficiency of 95.45%. From the
foregoing it evident that the Moringa oleifera seed from Adamawa State had the fastest turbidity
removal rate of 18NTU/minute within the first 30minutes. This is faster than 12.4NTU/minute and
15.53NTU/minute for Moringa oleifera samples from Yobe and Borno states respectively within
the same time interval. This result is in consonance with [30] which showed that locations of
plants species and their environments have effects on their chemical constituents. Additionally
[31] reported a difference in antioxidant activity of leaf extracts of Annona senegalensis between
leaves of Togo and Burkina Faso origin.
Furthermore [32] stated that seeds from different sources (geographic locations) exhibit
varying coagulation performance. This may have to do with differences in the protein content and
development of the seed [18]. The coagulation mechanism of the Moringa oleifera seed powder
has been attributed to adsorption and charge neutralization [33,34]; interparticle bridging [35] and
enmeshment by net like structure [35]. The compounds accomplishing coagulation in water are
low cationic peptides with molecular weight polyelectrolytes ranging in molecular mass of 616KDa with an isoelectric point (PI) value of around 10 and high molecular mass protein
component with molecular weight of 66KDa [36]. The small sized proteins of low molecular weight
accomplish coagulation by adsorption and change neutralization while flocculation by interparticle bridging is mainly characteristic of high molecular weight polyelectrolytes [18]. A large
molecular mass protein (66KDa) having coagulation activity was isolated from the seeds of
Moringa oleifera comparable with that of cationic peptides and alum but without antimicrobial
properties[37].

colour removal efficiency(%)

105
100
95
90
percentage
rem(adamawa)
percentage rem(borno)

85
80
75
0

200

400

600

800

1000

1200
1400
Contact time(mins)

1600

Fig. 5: Effect of contact time on colour removal

Consequently, Agrawal et al. [37] opined that the flocculation coagulation activity on Moringa
oleifera is a cumulative effect of different active components of the seed rather than isolated
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Nwaiwu N.E., Ibrahim W.I. and Raufu I.A., 2012. Antiseptic and Coagulation Properties of Crude Extracts of
Moringa Oleifera Seeds from North East of Nigeria.
.

constituents. Figure 5 shows the reduction of colour with time for the three Moringa oleifera seed
powder samples. Colour removal efficiencies of between83.66% - 98.32% was achieved with the
first 30mins with the sample from Adamawa state having the highest value of 98.32% at rate of
525pt/co per minute.
CONCLUSION
The Moringa oleifera seeds from Borno, Yobe and Adamawa states showed similar
antibacterial properties but different coagulation properties. The seeds from Adamawa state
exhibited the fastest turbidity and colour removal potential. Consequently it is being
recommended for large scale water treatment use in the North East region of Nigeria.
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Moringa Oleifera Seeds from North East of Nigeria.
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