ASSESSMENT OF ANTI-BACTERIA ACTIVITIES OF Moringa oleifera LEAF ON

DIFFERENT DRYING METHOD

By
BUKAR Amina Grema
NAS/BIO/18/

BEING A RESEARCH PROJECT SUBMITTED TO DEPARTMENT OF BIOLOGICAL

SICENCES, COLLEDGE OF NATURAL AND APPLIED SCIENCES AL-QALAM UNIVERSITY
KATSINA, IN PARTIAL FUFILLMENT FOR THE AWARD OF BACHELOR DEGREE Bsc. IN
BIOLOGY

JANUARY, 2023.
 Abstract
The antibacterial activity of Moringa oleifera Leaves extract belonging to the family Moringaceae, was
determined using agar well diffusion method against some selected bacteria. Methods: Mueller Hinton
Agar (MHA) (Becton Dicknson M. D USA), media was prepared according to the manufacturer’s
instruction. Sterile Mueller Hinton agar plates were inoculated with the test culture by surface spreading
using sterile wire loops and each bacterium evenly spread on the entire surface of the plate to obtain
uniformity of the inoculum. Concentrations of 30, 60, 90 and 120mg/ml prepared from the dry leaves
powder were used for antibacterial analysis using agar well incorporation methods. Plates of nutrents ager
were prepared and allowed to solidify on Petri dishes. Each plate was then seeded with a test bacterium.
Four holes were made in each of the plate with a sterile 2.0 mm diameter cork borers. Each of the four
holes was filled with a given concentration of the extract mixed with plane sterile agar. The plates were
then incubated at 37°c for 24 hours. The diameters of zones of inhibition were measured using a meter rule
and the mean value for each organism was recorded. The result shows that, the aqueous and ethanol
extracts of the plant leaves show an inhibitory effect on the growth of the tested bacteria. For aqueous and
ethanol extracts, the inhibitory effect on Escherichia coli was significantly higher (P<0.05) than that of
Staphylococcus aureus and Pseudomonas aeruginosa respectively. In addition, both ethanol and aqueous
extract showed a significantly higher (P<0.05) inhibitory effect at higher concentration of 120mg/ml.
Introduction

Moringa Oleifera has been used extensively in traditional medicine for the treatment of several ailments,

promotes digestion, skin diseases, diarrhea, as stimulant in paralytic afflictions, epilepsy and hysteria

(Farooq et al., 2012; Mishra et al, 2011). Various parts of the plant have been shown to be useful, such

as the roots have been experimentally shown to have anti-inflammatory action (Odebiyi and Sofowora,

1999; Ndiaye et al., 2002; Jamil et al., 2007), the leaves, stem bark and seeds have been reported to

have therapeutic properties (Anwar and Rashid, 2007).

 Aim and Objective of the Study

Aim of the study

The aim of this study is to asses the antibacterial activities of Moringa oleifera of different drying

method.

Objectives of the Study

1. To determine the phytochemical component of Moringa oleifera.

2. To determine the antibacterial susceptibity of Moringa oleifera.

 Material and methods
MATERIALS GLASS WARES
EQUIPMENT  Conical flask
 Mortar and pestle  Test tubes
 Filter paper  Glass rod
 Blade CHEMICAL/ REAGENTS
 Incubator  Distilled water
 Hot plate  Safranin
 Water bath  Ethanol
 Autoclaving machine  3.1.4 MEDIA
 Lamina safety  Nutrient Agar
 Incubator shaker  Nutrient broth
 Muslim cloth
 Filter paper
Method of sample collection

The leaves of Moringa oleifera was collected from three different areas in Katsina

metropolis. The areas were Fatima Shema Estate, Tudun Matawalle and Kofar Kwaya

Opposite Dahiru Barau Mangal’s residence.

Sample preparation Sorting
Fresh, green, undamaged, non-insect infected were selected. Bruised,
discoloured, decayed and wilted leaves were discarded before washing the
leaves.
 Washing
 Drying techniques
 Sun drying
 Shade drying
 Oven drying
 Aqueous
 Ethanol
3.6 Chemical analysis
The dried leaves was analysed for proximate composition (protein, fat, fibre,
carbohydrate and ash), vitamins (vitamin B1, vitamin A), minerals (iron,
iodine, zinc, calcium) and anti-nutrients (phytate, oxalate, saponin, tannin).
PHYTOCHEMICAL SCREENING
 Tannins

 Saponins

 Flavonoids

 Terpenoids

 Glycosides

 Alkaloids

 Phenolic Compounds

Sensitivity Test

 Preparation of Media

 Culture
INCLUDE REFERENCES
 Disc Difusion
 RESULT
Table 4.1 shows the phytochemical screening of Moringa oleifera leaf of ethanol extract
Sample Alkaloids Saponins Flavonoid Glycoside Tannis Terpenoids Phenolic
Coumpound

Sundry + - - + + - -

Ovendry ++ + - + ++ - -

Shade dry +++ +++ - ++ +++ - +++

Key: + = indicate presence and - = indicate absence

THIS IS NOT A SCIENTIFIC TABLE

Table 4.2 shows the phytochemical screening of Moringa oleifera leaf of Aqueous extract
Sample Alkaloids Saponins Flavonoid Glycoside Tannis Terpenoids Phenolic
Coumpound

Sundry + - - + + + +

Ovendry ++ ++ - ++ ++ ++ ++

Shade dry +++ - - +++ +++ - -

Key: + = indicate presence and - = indicate absence

Table 4.3 antibacterial susceptibility profiles of moringa oleifera leafs of staphylococcus, E coli
and seudomonas,
Bacteria ACOOL ECOOL ACSOL ECSOL ACSHOL ECSHOL ACOOL ECOOL ACSOL ECSOL ACSHOL ECSHOL +C
+C +C +C +C +C
  Conc. (mg/ Conc. (mg/ Conc. (mg/ Conc. (mg/ Conc. (mg/ Conc. (mg/            
ml) ml) ml) ml) ml) ml)

Staphylococcus
100 Nil 100 20 100 20 100 22 100 2 100 2 25 26 24 22 9 22
75 Nil 75 8 75 10 75 17 75 2 75 2 25 26 24 22 9 22
50 Nil 50 Nil 50 Nil 50 15 50 2 50 2 25 26 24 22 9 22
25 14 25 5 25 Nil 25 13 25 2 25 8 25 26 24 22 9 22

 
                                   
E. Coli 100 10 100 2 100 12 100 19 100 2 100 12 31 29 28 28 20 27
75 Nil 75 2 75 Nil 75 13 75 2 75 9 31 29 28 28 20 27
50 5 50 Nil 50 2 50 8 50 2 50 Nil 31 29 28 28 20 27
25 3 25 12 25 Nil 25 2 25 Nil 25 Nil 31 29 28 28 20 27
 
                                   
Seudomonas
100 Nil 100 Nil 100 Nil 100 Nil 100 Nil 100 Nil 24 24 28 30 23 22
75 Nil 75 Nil 75 Nil 75 Nil 75 Nil 75 Nil 24 24 28 30 23 22
50 Nil 50 Nil 50 Nil 50 Nil 50 Nil 50 Nil 24 24 28 30 23 22
25 Nil 25 Nil 25 Nil 25 Nil 25 Nil 25 Nil 24 24 28 30 23 22

Keys: Conc.=Concentration, O=Oven Dry, S=Sun Dry, Sh.= Shade Dry, +Control= Positive control, E.
coli= Escherichia Coli.

THIS IS NOT A SCIENTIFIC TABLE

Discussion of Findings

The rising prevalence of pathogenic microorganisms' resistant to the newer antibiotics has been

expressed in the last three decades (Valarmathy et al., 2010). The current study exhibits that Moringa oleifera

leave extract shows phytochemical bioactive compounds like flavonoids, tannins, alkaloids, saponins and

phenols compound in aqueous and ethanol extract shows in Table 4.1 and 4.2 respectively. That they exhibit

antibacterial activity.

Alkaloids are natural bioactive compounds contain basic nitrogen atoms. They also have

pharmacological effect and are used as herbal medications (Rhoades, 1979). Flavonoids promote the effect of

vitamin C and act as antioxidants. They are also known to be biologically active against liver toxins, tumors,

viruses and other microbes (Korkina, 1997). Tannins have revealed potential Antiviral, Antibacterial and

Anti-parasitic effects. Saponins cause hemolysis of red blood cells (Winter, 1993).
 Antibacterial activity of Moringa oleifera was seen against several bacteria namely Escherichia Coli,

Pseudomonas aeruginosa, Staphylococcus aureus, Proteus vulgaris, Streptococcus mutants, Bacillus subtilus

and Staphylococcus epidermidis (Naplean 2009). The ethanol extract showed antibacterial effect against three

different bacteria Staphylococcus, E. coli and Seudomonas (Table 4.3). The results showed that increasing the

concentration of the Aqueous extract and ethanol extracts increased the zone of inhibition. The medicinal plant

Moringa oleifera exhibits good antibacterial activity against, Staphylococcus, and E. coli in this study.

However, many previous results revealed the strains E. coli, P. aeruginosa and S. enterritidis were resistant to

many treatments (Jackson et al., 2011). This resistance observed were matched from a study on the

antibacterial properties of Indian plants showing Moringa extracts to be ineffective against E. coli. Thus, the

results in current study revealed significant inhibitory effect of Aqueous and ethanol leaves extract of Moringa

oleifera on the Staphylococcus and E. coli.

Conclusion

This study has revealed that the Aqueous and ethanol extracts of Moriga oleifera leaf possess some degree of
antibacterial effects. Were the leaf extracts shown to contain bioactive compounds with a clear antibacterial
activity, capable of inhibiting the growth of gram negative bacteria, E. coli and Staphylococcus. More and
further studies can achieve that Moringa oleifera can be used to discover which bioactive compounds
responsible on the antibacterial activity.

Recommendations

The researcher recommend the following points based on the study.

i. There is also the need for more work to be carried out to establish the conflicting findings from various
laboratories on the efficacy of the leaf extracts on different bacteria’s for medical and herbal uses.
ii. Moringa oleifera represents an economic and safe alternative to treat infectious diseases in addition to its
many other uses.
iii. Its use is thus highly encouraged and more work should be carried out to determine the antibacterial
constituents that can be used for drug formulation.
Some of the references
Anwar, E, Rashid, U. (2007). Physico-chemical characteristics of Moringa oleifera seeds and seed oil from a
wild provenance of Pakistan. Pakistan Journal of Botany, 39: (5) : 1443-1453.
Farooq, E, Rai, M., Tiwari, V., Khan, A. A., Farooq, S. (2012). Medicinal properties of Moringa
oleifera: An overview of promising healer. Journal of Medicinal Plants Research, 6: (27): 4368-
4374.
Jamil, A., M., Shahid, M. M., Khan, M., Ashraf, (2007). Screening of some medicinal plants for
isolation of antifungal proteins and peptides. Pakistan Journal of Botany, 39: (l): 211-221.
Mishra, G, Singh, P. , Verma, R. , Kumar, S., Srivastav, S., Jha, K.K., Khosa, R. L. (2011).Traditional
uses, Phytochemistry and Pharmacological properties of Moringa oleifera plant: An overview.
Der Pharmacia Lettre, 3: (2): 141164.
Odebiyi, A., Sofowora, E. A. (1999). Pytochemical screenings of Nigerian medicinal plants part ll.
Lyodia, 44 : 234-246

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