Phyto-Chemical Screening and Antibacterial Activities of Aqueous Extracts of Diospyros mespiliformis, Mitragyna inermis, Piliostigma reticulatum and Tamarindus indica in Northern Nigeria
The phytochemical contents and antimicrobial activities of leaves and root bark extracts of Diospyros mespiliformis, Mitragyna inermis, Piliostigma reticulatum and Tamarindus indica were investigated. The plant materials were collected along River Kubwa in Damboa local Government area of Borno State, Nigeria. The crude extracts of these plant materials were prepared based on soxhlet method using soxhlet extractor. About 400mg of each pulverized sample was used for the extraction process. Three (3) liters of distilled water was used for the extraction process. All the extracts showed the presence of carbohydrates, cardiac glocosides, saponins, tannins and flavonoids. Terpenoids was detected in Diospyros mespiliformis, Tamarindus indica extracts and Piliostigma reticulatum root bark extract. Disc sensitivity technique was conducted on eight bacteria isolates. These bacteria are: staphylococcus aureus, bacillus subtilis, Escherichia coli, salmonella typhi streptococcus faecolis pseudomonas aeruginosa coreynebacteria species and shigelladysenteriae. The growth inhibition studies on the test isolates indicated that most of the plant extracts had significant effects against some of the isolates (microorganisms). The antimicrobial activities were subjected to Two-way Analysis of Variance (ANOVA) and it indicated that there was statistically (P < 0.05) significant difference at different concentrations. It is concluded that these plant materials have the potential of improving the bacteriological quality of pond or surface water (which most rural communities in developing countries depend for many purposes) that are heavily polluted with pathogenic microbes.
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Phyto-Chemical Screening and Antibacterial Activities of Aqueous Extracts of Diospyros mespiliformis, Mitragyna inermis, Piliostigma reticulatum and Tamarindus indica in Northern Nigeria
2. Arid Zone Journal of Engineering, Technology and Environment, April, 2017; Vol. 13(3):420-427
ISSN 1596-2490; e-ISSN 2545-5818; www.azojete.com.ng
421
2003). This is possible if traditional water sources can be treated using water purification
methods, which are inexpensive and suitable under local conditions. One area that holds a lot of
prospect for the future is the plant kingdom and particularly the use of these plant seeds, leaves,
root barks, stem barks and fruits as natural coagulants to remove turbidity and bacteria in surface
water (Ebobganya et al., 2013, Ghebremichael et al., 2005). It has also been observed by many
researchers that reduction in turbidity is associated with significant improvements in
bacteriological quality (Ghebremichael et al., 2006; Marobhe, 2008b; Conte et al., 2007; Phani
and Rajkumar, 2013).
The present study examined the performance of natural plant materials that are locally available
in North East Nigeria as renewable sources of natural coagulants using their leaves and root
barks aqueous extracts in pond water treatment.
2. Materials and Methods
2.1 Collection of Plant Materials
The fresh root barks and matured leaves of Diospyros mespiliformis, Mitragyna inermis,
Piliostigma reticulatum and Tamarindus indica were collected from bush along Kubwa riverside
in Damboa local Government area of Borno State, Nigeria in the month of April, 2013, when the
leaves of these trees were green. The root barks and matured leaves of these plants were
removed, chopped into pieces using mortar and pestle and shade dried at room temperature. Each
dried sample was subjected to a size reduction using mechanical grinding machine.
2.2 Preparation of Plant Extracts
The extraction process was conducted based on soxhlet method using soxhlet solvent extractor.
About 400mg of each pulverized sample was used for the extraction process. Three liters of
distilled water was added to cover the sample into solution. The heating mantle on the extractor
was switched on and allowed to reflux the mixture for 2hrs. The solution was removed and
filtered to remove debris. The aqueous extracts were evaporated to dryness on rotary evaporator.
2.3 Source of Raw Water
The raw water having turbidity strength of 422 NTU was collected from a pond along Molai
Road, Maiduguri, Nigeria. Maiduguri the capital of Borno State lies within the latitude 11°
51’ N
and 13°
40’ N and longitude 10°
0’ E and 13°
E. The water sample was collected towards the
middle of the pond to serve as representative sample of the whole pond water and a laboratory
test was carried out each day the sample was collected.
2.4 Coagulation Tests
The coagulation activities of the aqueous crude root bark and leave extracts of the plant materials
on highly turbid water (422 NTU) were evaluated using an experimental jar tester of Phipps and
Birds fitted with six (1000ml) beakers. The coagulation tests (jar test experiments) were carried
out at varying dosages of the aqueous crude extracts using standard procedure described by
Gurchran and Jagdish (2009).
2.5 Statistical Analysis
3. Yadima et al.: Turbidity and colour reduction of pond water using extracts of diospyros mespiliformis,
mitragyna inermis, piliostigma reticulatum and tamarindus indica in northern Nigeria. AZOJETE,
13(3):420-427. ISSN 1596-2490; e-ISSN 2545-5818, www.azojete.com.ng
422
T-Test analysis was used to compare the significant differences of turbidity and colour removals
of the aqueous leaves and root bark extracts of the plant materials on the raw water
3. Results and Discussions
The Percentage of turbidity and colour removal using aqueous root bark extracts of the plants are
shown in table 1.
Table 1: Percentage of turbidity and colour removal using aqueous root bark extracts
Number of
Jars
Concentrations of
extracts (mg/l)
Settling
time (min)
Plant
extracts
Percent turbidity
removal (%)
Percent colour
removal (%)
J1 60 20
DR 9.00 8.71
MR 8.53 8.25
PR 0.95 1.24
MR 10.24 12.35
J2 75 23
DR 9.48 9.17
MR 8.77 8.48
PR 1.9 2.25
MR 12.38 14.92
J3 90 24
DR 9.95 9.62
MR 10.09 9.77
PR 2.38 3.19
MR 11.9 14.64
J4 105 26
DR 9.72 9.4
MR 9.86 9.54
PR 7.62 8.03
MR 14.76 17.79
J5 120 28
DR 8.53 8.25
MR 7.82 7.57
PR 2.62 3.99
MR 10.71 12.92
J6 135 30
DR 14.22 13.76
MR 10.31 9.97
PR 2.14 2.59
MR 11.67 14.34
Key: DR = Diospyros mespiliformis root bark MR = Mitragyna inermis root bark PR = Piliostigma reticulatum root bark TR=Tamarindus indica
root bark
The percentage of turbidity and colour removal using root bark extracts of the plant materials are
presented in Table 1. From the Table, it can be seen that the highest turbidity removal of
Diosppyros mespiliformis extract was 14.22% at the dosage of 135mg/l, for Mitragyna inermis, it
was 10.31% at a dose concentration of 135mg/l, for Piliostigma reticulatum, it was 7.62% at a
dosage of 105mg/l and for Tamarindus indica, it was 14.76% at a dose concentration of 105mg/l.
Diosppyros mespiliformis had colour removal efficiency in the range of 8.25% to 13.76% and the
highest removal was achieved at dose concentration of 135mg/l. For mitragyna inermis extract,
is from 7.54% to 9.97% and had the highest colour removal ability at the dose concentration of
135mg/l. The percentage of colour removal for Piliostigma reticulatum and Tamarindus indica
4. Arid Zone Journal of Engineering, Technology and Environment, April, 2017; Vol. 13(3):420-427
ISSN 1596-2490; e-ISSN 2545-5818; www.azojete.com.ng
423
ranged from 1.24% to 8.03% and 12.35% to 17.79% respectively and the highest efficiencies
were achieved at dose concentration of 105mg/l for the extracts.
The Percentage of turbidity and colour removal using aqueous leaves bark extracts of the plants
are shown in table 2.
Table 2: Percentage of turbidity and colour removal using aqueous leaves extracts
Number
of Jars
Concentrations of
extracts (mg/l)
Settling time
(min)
Plant extracts Percent turbidity
removal (%)
Percent colour
removal (%)
J1 60 20
DL 9.00 8.71
ML 8.77 8.64
PL 0.71 1.17
TL 14.05 16.94
J2 75 23
DL 10.19 9.86
ML 9.24 8.94
PL 3.1 5.1
TL 15.48 18.66
J3 90 24
DL 9.9 9.58
ML 8.59 8.58
PL 5.24 6.22
TL 15 18.09
J4 105 26
DL 9.72 9.4
ML 10.19 9.86
PL 2.62 4.31
TL 14.29 17.21
J5 120 28
DL 9.48 9.17
ML 8.91 8.62
PL 2.38 3.92
TL 17.14 20.66
J6 135 30
DL 11.02 10.66
ML 8.53 8.23
PL 3.81 6.27
TL 18.57 22.38
Key: DL=Diospyros mespiliformis leave ML=Mitragyna inermis leave PL=Piliostigma reticulatum leave
TL=Tamarindus indica leave
The percentage of turbidity and colour removal using leave extracts of the plant materials are
presented in Table 2. The percentage of turbidity removal efficiency by Diosppyros
mespiliformis leaves extract varied from 9.00% to 11.02%, for mitragyna inermis, it varied from
8.53% to 10.19%, for Piliostigma reticulatum, it varied from 0.71% to 5.14% and for
Tamarindus indica, it varied from 14.05% to 18.57%. The highest percentage turbidity removal
for Diosppyros mespiliformis, Mitragyna inermis, Piliostigma reticulatum and Tamarindus
indica were achieved at dose concentrations of 135mg/l, 105mg/l, 90mg/l and 135mg/l
respectively.
The percentage of colour removal using the leave extracts (table 2) indicated that for Diosppyros
mespiliformis, it varied from 6.71% to 10.66%, for Mitragyna inermis, it was from 8.23% to
9.86% with the lowest percentage removal of 8.23% at the dosage of 135mg/l. Piliostigma
5. Yadima et al.: Turbidity and colour reduction of pond water using extracts of diospyros mespiliformis,
mitragyna inermis, piliostigma reticulatum and tamarindus indica in northern Nigeria. AZOJETE,
13(3):420-427. ISSN 1596-2490; e-ISSN 2545-5818, www.azojete.com.ng
424
reticulatum extract had percentage of colour removal which varied from 1.17% to 6.27% and
Tamarindus indica extract had percentage removal of colour which varied from 16.94% to
22.38%.
TheT-Test analysis for the coagulation property (turbidity removal) of the aqueous root bark and
leaves extracts of the plants are shown in table 3
Table 3: T-Test analysis for the coagulation property (turbidity removal) of the aqueous root
bark and leaves extracts
Diospyros mespiliformis root extract Diospyros mespiliformis leave extract
N Mean StDev T Df P-value Decision N Mean StDev T Df P-value Decision
IT 6 422.00 0.00 12.09 5 0.000 Reject Ho IT 6 422.00 0.00 35.30 5 0.000 Reject. Ho
FT 6 397.17 8.68 FT 6 380.29 2.89
Mitragyna inermis root bark extract Mitragyna inermis leave extract
N Mean StDev T Df P-value Decision N Mean StDevT T Df P-value Decision
IT
FT
6
6
422.00
383.05
0.00
4.22
22.62 5 0.000 Reject Ho IT
FT
6
6
422.00
383.07
0.00
2.62
35.72 5 0.000 Reject Ho
Piliostigma reticulatum root bark extract Piliostigma reticulatum leave extract
N Mean StDev T Df P-value Decision N Mean StDev T Df P-value Decision
IT 6 420.00 0.00 3.04 5 0.029 Reject Ho IT 6 422.00 0.00 4.82 5 0.005 Reject Ho
FT 6 407.67 9.93 FT 6 407.50 6.35
Tamarindus indica root bark extract Tamarindus indica leave extract
N Mean StDev T Df P-value Decision N Mean StDev T Df P-value Decision
IT 6 420.00 0.00 18.41 5 0.000 Reject Ho IT 6 420.00 0.00 21.86 5 0.000 Reject Ho
FT 6 369.83 6.68 FT 6 353.83 7.41
Ho = There is no difference in turbidity removal of the extracts in the raw water
The Table indicated that the pre-determined significance values of all the extracts were less than
0.05.
The coagulation activities of the plant extracts were found to vary slightly at various dosages.
The highest percentage of turbidity and colour removal of 18.57 and 22.38 was demonstrated by
Tamarindus indica leaves extract while the lowest percentage removal of turbidity and colour of
0.71 and 1.17 respectively was demonstrated by Piliostigma reticulatum leaves extract.
Generally, it was observed that the coagulating effect of these plant materials were not
substantial. The patterns of floc-formation for all the plant extracts were observed to be of
smoky-like than aggregating into sizes that may have enough density to settle down in a short
period of time. This might be attributed to some factors such as the initial pH of the raw water
which stood at a value of 8.1.
The results of T-Test analysis for the coagulation property (colour removal) of the aqueous root
bark and leaves extracts of the plants are shown in table 4.
6. Arid Zone Journal of Engineering, Technology and Environment, April, 2017; Vol. 13(3):420-427
ISSN 1596-2490; e-ISSN 2545-5818; www.azojete.com.ng
425
Table 4: T-Test analysis for the coagulation property (colour removal) of the aqueous root bark
and leaves extracts
Diospyros mespiliformis root bark extract Diospyros mespiliformis leave extract
N Mean StDev T Df P-value Decision N Mean StDev T Df P-value Decision
IC 6 4361.00 0,0 12.09 5 0.000 Reject Ho IC 6 4361.00 0.0 35.34 5 0.000 Reject Ho
FC 6 3932.7 86.8 FC 6 3943.8 289
Mitragyna inermis root bark extract MItragyna inermis leave extract
N Mean StDev T Df P-value Decision N Mean StDev T Df P-value Decision
IC 6 4361.00 0.0 22.62 5 0.000 Reject Ho IC 6 4361.00 0.0 37.32 5 0.000 Reject Ho
FC 6 3971.5 42.2 FC 6 3978.0 25.1
Piliostigma reticulatum root bark extract Piliostigma reticulatum leave extract
N Mean StDev T Df P-value Decision N Mean StDev T Df P-value Decision
IC 6 4357.00 0.0 3.66 5 0.015 Reject Ho IC 6 4357.00 0.0 5.83 5 0.002 Reject Ho
FC 6 4202.2 103.6 FC 6 4161.0 82.4
Tamarindus indica root bark extract Tamarindus indica leave extract
N Mean StDev T Df P-value Decision N Mean StDev T Df P-value Decision
IC 6 4357.00 0.0 18.63 5 0.000 Reject Ho IC 6 4357.00 0.0 21.90 5 0.000 Reject Ho
FC 6 3725.5 83.0 FC 6 3529.7 92.5
Ho = There is no difference in colour removal of the extracts in the raw water
The Table indicated that the pre-determined significance values of all the extracts were less than
0.05.
It was often reported that the performance of natural coagulants improves at acidic pH of raw
water samples (Bodlund, 2013). Similarly, Marobhe, (2008b) observed that the net surface
charge of colloidal particles is reduced at low pH and hense the electrostatic repulsion between
the colloids and the thickness of the double layer is also reduced. This suggests that efficient
particles destabilization and optimal flocculation occur at lower pH. The floc-formation patterns
of these extracts could also be explained by the degree of their solubility or polarity in the water
(solvent). The best performance of a crude extract from a finely grinded seed powder could be
due to its large total surface area; whereby most of the water soluble proteins are at the solid-
liquid interface during the extraction process and this might increase the concentration of active
coagulation polymer in the extracts which improves the coagulation process (Hart, 2000).
Therefore, these extracts require substantial settling time before a significant turbidity and colour
removal can be achieved, hence they could be used with coagulant aid such as Non-ionic
polymer which may raise the pH as well as increase the alkalinity. The T-Test results of turbidity
and colour removal abilities of the plant extracts indicated that the pre-determined significance
values of all the extracts were less than 0.05. This signified that there were differences in the
removal of turbidity and colour at the varying dosages of the extracts.
7. Yadima et al.: Turbidity and colour reduction of pond water using extracts of diospyros mespiliformis,
mitragyna inermis, piliostigma reticulatum and tamarindus indica in northern Nigeria. AZOJETE,
13(3):420-427. ISSN 1596-2490; e-ISSN 2545-5818, www.azojete.com.ng
426
4. Conclusion
The highest percentage of performance for turbidity removal was 18.57% and for colour removal
was 22.38% and was established by Tamarindus indica leaves extract which was rather low. The
implication was that these plant materials could be used with the help of an aid (coagulation aid)
to improve their performances.
This investigation has opened up the possibility of using these plant materials as natural
coagulants in improving the quality of pond or surface water (on which most rural communities
in developing countries depend for drinking and other purposes) that are highly turbid.
Therefore, it is recommended that the use of these plant materials should be encouraged among
rural communities to improve the quality of pond or surface water. Toxicity test should be
carried out on these plant extracts so that safe application concentrations can then be determined,
and further diversification of these plant extracts towards treating some common chemicals
including heavy metals and pathogenic organisms and other related contaminants in pond or
surface water is also recommended.
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