Republic of the Philippines

DEPARTMENT OF EDUCATION
Region I
Schools Division Office I Pangasinan
Mapandan National High School
Mapandan, Pangasinan

Antibacterial Activity of Malabar Nightshade

(Basella alba) Leaf Ethanolic Extract

Against E. coli (Escherichia coli)

In partial fulfillment

of the requirements in Research II

Juan Lorenzo N. Soriano

Mar Clarence DV. Quinto
Llanah Roubbie M. Quiray
Researchers

Jean Leah J. Mangapot

Research Adviser
 ACKNOWLEDGEMENT

The researchers’ sincerest appreciation and warmest thanks are extended to all

individuals who have given their contributions to the success of this study:

To Ma’am Jean Leah J. Mangapot, Master Teacher II, Science Department as

their research adviser, who gave her support, guidance and assistance throughout the

preparation of this study.

To the Head Teacher of the Science Department, Madam Marie Joy T. Visperas,

who gave her full support and guidance throughout the study.

To the Principal of Mapandan National High School, Madam Elvira O. Velasquez

who encouraged and gave moral support for the success of the Research work.

To Sir Ces A. Ramirez, who is always there to support and for sharing his

knowledge.

To the Virgen Milagrosa University Foundation and Lyceum Northwestern

University as the testing laboratory.

To their parents who have given inspiration and moral support throughout this

endeavor.

Above all to almighty GOD whom the researchers owe their success for providing

wisdom, fresh supply of grace, treasures to enrich their soul, and guidance to have an

excellent choice and a ripples blessing in effect.

 The Researchers

DEDICATION

This piece of work is whole heartedly dedicated to the researchers’ family who

gave their moral support in the making of this valuable work, thus, giving them courage

whenever they looked down and lose hope. They are the one who cheered and uplifted

them to accomplish this work.

To their principal, head teacher, and teachers for sharing their professional

expertise and ideas to conduct their study.

To their friends who are there along the way just to support and make them

inspired.

Above all, to our almighty God, for His countless blessings, giving the, strength,

courage, knowledge, and wisdom in conducting this study.

To all of you, they owe you a lot.

The Researchers
 ABSTRACT

“Antibacterial Activity of Malabar Nightshade (Basella alba) Leaf Ethanolic

Extract against E. coli (Escherichia coli)”

Soriano, Juan Lorenzo N.; Quinto, Mar Clarence DV.;Quiray, LlanahRoubbie M.

Grade IX-SSC Students, Mapandan National High School, Mapandan, Pangasinan.

The researchers devolved on the study of utilization of antibacterial property of

Malabar Nightshade leaf ethanolic extract against E. coli.

Generally, this study aimed to test the antibacterial activity of Malabar Nightshade

leaf ethanolic extract against E. coli. Specifically, it sought to find phytochemicals

present in Malabar Nightshade leaf ethanolic extract. Also to find which concentration of

Malabar Nightshade leaf ethanolic extract is the most susceptible against E. coli and its

significant difference to the positive control.

The Malabar Nightshade leaves were gathered, washed, cut and weighed. The

leaves were subjected to maceration, extraction and Phytochemical screening. The results

show that it contains alkaloidal salt, saponin, glycosides and non-hydrolizable tannins.

After the test was done, four different concentrations were prepared for the study.

Using Disc Diffusion Method, the different concentrations in Mueller Hinton agar plate

with inoculums of E. coli was tested. The effect of Malabar Nightshade leaf ethanolic

extract was tested using the Completely Randomized Design (CRD) in which a group of
test materials will be studied only but subsequent treatment is applied to determine the

antibacterial property of Malabar Nightshade leaf ethanolic extract against E. coli.

The study was able to prove that the presence of alkaloidal salt certifies that the

Malabar Nightshade leaf ethanolic extract has antibacterial property. The 100% and 75%

concentration of Malabar Nightshade leaf ethanolic extract with 80 % susceptibility of its

antibacterial property yields best among the treatment used. The positive control has 90%

susceptibility while the antibacterial property of Malabar Nightshade leaf ethanolic

extract has only 80% susceptibility.

Based on the results of the study, the used of Malabar Nightshade leaves are

highly recommended to a person suffering from disease or infection caused by E. coli

bacteria. It is also cheaper, safer and economical than the commercial anti-bacteria found

in the market.
 CHAPTER I

INTRODUCTION

A. Rationale and Background of the Study

Plants are said to be the backbone of all life on Earth and an essential resource for

human well-being. As a result, it is widely use as medicine. However, a considerable

number of people in our generation still do not recognize the value and significance of

medicinal plants, many people continue to rely on synthetic drugs for treatment of

common illnesses irrespective of the expensive price they are been sold.

Bacteria, a large domain of prokaryotic microorganisms, is the most common and

important causative agent of diseases among the microorganisms. It can be beneficial as

well as pathogenic to human health. Numerous of bacterial cells live in the human flora

most especially in the gut and skin flora. Bacteria have the great challenging capacities to

our health and may cause cells, tissues, organs or organ systems seriously damaged. It

can cause from simple infections to fatal or life threatening infections in the living things

including human beings.

Gram negative bacteria are major resistant to antibiotics. It is spread worldwide,

in virtually all environments that support life. It make-up a small portion of skin flora,

gastrointestinal tract flora and different more floras in normal human body. But it also

includes pathogenic bacteria. Several of its species can cause many types of infections
and diseases like foodborne and waterborne illnesses. Because of its resistance to

antibiotics, it is difficult to treat that is why many drugs are ineffective to its several

species. Also, some drugs have toxic side effects.

Many bacterial isolates are cause by contaminated food, contaminated water or

even contaminated environment. People should assure that the foods they are eating are

safe and free from pathogenic bacteria. They should avoid foods from streets or foods

that came from uncertain areas. If they are infected, they should be careful on the

different drugs or anti-bacteria that they will be using. They should use anti-bacteria that

is economically friendly so that they can assure that it doesn’t have side effects. In other

side, they can also help our environment.

This study aimed to test the Antibacterial Activity of Malabar Nightshade leaf

ethanolic extract against E. coli. To find better way of preventing diseases cause by E.

coli that is safer and economical for the community. Also, for the community to be aware

of the health benefits of Malabar Nightshade leaves.

B. Statement of the Problem

The study aimed to test the antibacterial property of Malabar Nightshade leaf

ethanolic extract against E. coli.

Specifically, it sought to answer the following questions:

1. What are the phytochemicals present in Malabar Nightshade leaf ethanolic

extract?

2. Which among the treatment with varying concentrations (100%, 75%, 50% and

25%) yields the highest zone of inhibition?

 3. Is there any significant difference between the antibacterial property of Malabar

Nightshade leaf ethanolic extract and the positive control?

C. Hypotheses

H0 – There is no significant difference between the antibacterial property of

Malabar Nightshade leaf ethanolic extract and the positive control against E. coli.

H1 – There is a significant difference between the antibacterial property of

Malabar Nightshade leaf ethanolic extract and the positive control against E. coli.

D. Significance of the Study

This study will be beneficial to the following:

To the pharmaceutical companies in formulating drugs against diseases and

infections cause by E. coli from the Malabar Nightshade leaf ethanolic extract. The

results of the study may lead to the discovery of a more superior and economical

antibacterial product than the commercial one.

To the people of the community to be aware of the benefits of the plant and the

results will be utilized for the treatment of certain diseases and infections in the

community. This may improve the health care in the community, to meet a healthy

country.

To the future researchers in utilizing the knowledge gained from the study in

curbing the bacterial samples using the Malabar Nightshade leaf ethanolic extract.
E. Scope and Delimitations

The research study was conducted to find out the antibacterial activity of Malabar

Nightshade leaf ethanolic extract against E. coli. It sought to find phytochemicals present

in Malabar Nightshade leaf ethanolic extract. Also to find which concentrations of

Malabar Nightshade leaf ethanolic extract has the highest susceptibility against E. coli

and its significant difference to the positive control.

The study delimits on finding the efficiency of herbal medicine in regards and

comparison to modern medicines in treating diseases and infections caused by E.coli

which was tested at 0.05 level of significance. All analysis of the Malabar Nightshade

leaf ethanolic extract, synthetic antibiotics and bacterial strains needed for the completion

of the study were done in the month of August, 2016 at the Lyceum Northwestern

University - College of Pharmacy for phytochemical screening and the Microbiology

Laboratory of Virgen Milagrosa University Foundation - College of Medicine, San

Carlos City, Pangasinan for antibacterial assay.

Research Paradigm

Independent Variable Dependent Variable

1. Level of concentration Zones of inhibition of the

of Malabar bacterial sample against the
Nightshade leaf Malabar Nightshade mature
ethanolic extract at leaf ethanolic extract at
different different concentrations and
concentrations: positive control.
A. 100%
B. 75%
C. 50%
D. 25%

1. Test bacteria
A. Escherichia
coli

2. Positive Control
A. Ciprofloxacin

Intervening Variable

i. pH

ii. Temperature

Figure 1: The Research Paradigm of the Study

Figure 1shows the research paradigm of the study and illustrates that the study

will utilize the independent, dependent and intervening variables. The independent

variable shows that the study will be tested on both experimental and control basis where

the different level of concentration of 100%,75%,50% and 25% from the Malabar
Nightshade mature leaf ethanolicextract will be used to test the antibacterial activity of

the bacterial samples and ciprofloxacin will serve as the positive control.

Research Framework

This study will help to establish the antibacterial activity of Malabar Nightshade

leaf ethanolic extract which can be useful to fight against diseases causing bacteria of

which scientists believe that it is the source of half of human disease.

Thus, microbes are responsible for more illness or disease which explains the

phenomenon of disease transmission and provide viable basis upon which to make

decision about how to prevent illness. Hence, antibacterial is an agent that interferes with

the growth and reproduction of bacteria which may either kill or inhibit the growth of

bacteria.

Therefore, the Malabar Nightshade leafethanolic extract will yield an antibacterial

agent which inhibit bacteria and at the same time suppress their growth and ability to

reproduce thereby showing the antibacterial activity.

F. Definition of Terms

Antibacterial Activity – refers to the effect of any substances or drugs that inhibits or

destroys the reproduction of bacteria. Prevention of the bacteria to grow.

ANOVA (Analysis of Variance) - analysis tool used in statistics that splits the

aggregate variability found inside a data set into two parts: systematic factors and random

factors. The systematic factors have a statistical influence on the given data set, but the

random factors do not. Analysts use the analysis of the variance test to determine the

result independent variables have on the dependent variable amid a regression study.

Disc Diffusion Method - most widely used method for determining antibacterial

resistance in private veterinary clinics.

E. coli – a gram negative bacteria that is commonly found in the lower intestine of warm-

blooded organisms; one of the harmful bacteria nowadays; used as bacteria in this

research.

Ethanol – the principal type of alcohol found in alcoholic beverages, produced by the

fermentation of sugars by yeasts.

Extract –to remove (something) by pulling it out or cutting it out

Extraction- the separation of medicinally active portion of Malabar Nightshade

leaf. The Malabar Nightshade leaves areseparated from the inactive or inert

components through the use of ethyl alcohol as selective solvent and a standard

extraction procedure.

Gram Negative Bacteria - group of bacteria that do not retain the crystal violet stain

used in the gram staining method of bacterial differentiation.

Malabar Nightshade - an Asiatic climbing plant (Basellaalba) with fleshy shining

leaves and small white racemes flowers that is grown in the tropics as a potherb

and in temperate regions as an ornamental vine. Plant material used in the study.

Phytochemical Analysis – refers to the laboratory investigation on the active

constituents of Malabar Nightshade leaves.

Zone of inhibition – This area on a Muller Hinton agar plate where growth of an

organism is prevented by an antibiotic usually placed on the agar surface.

 CHAPTER II

REVIEW OF RELATED LITERATURE AND STUDIES

Related Literature

This chapter presents comprehensive informants about Malabar Nightshade which

include Hierarchical Classification, Botanical Origin, Distribution, Adaptability and

Availability, Propagation, Uses, some literature about E. Coli Bacteria and some Related

Studies associated with antibacterial activity of the plant.

Hierarchical Classification

Kingdom: Plantae

Division: Basella

Class: Basellaceae

Order: Caryophyllales

Family: Basellaceae

Genus: Basella

Species: B. alba

Scientific name: Basellaalba

Common Names: Malabar Nightshade

Malabar Spinach

Alugbati
Botanical Origin

Malabar Nightshade is a widely cultivated, cool season vegetable with climbing

growth habit. It is a succulent, branched, smooth, twining herbaceous vine, several meters

in length. Stem are Purplish or green. Leaves are fresh, ovate or heart-shaped, 5 to 12 cm

long, stalked, tapering to a pointed tip. Spikes are auxiliary, solitary, 5-29 cm long, and

purple when mature. Mainly leaves and stems are used for the medicinal purpose (J.

Suguna et al. Ijppr.Human, 2015; Vol. 3 (2): 66-77.)

Distribution

Malabar Nightshade is usually considered as native plant of Southern Asia

(India). In Southeast Asia and China, it has been grown since ancient times. It is now

widely cultivated in tropical Asia and America, and is given grown in temperate zones as

an annual in Southeast Asia and particularly popular in Malaysia and the Philippines. (M.

Rahmansyah 2016)

Bacteria

E. coli (Escherichia coli)

E. coli bacteria were discovered in the human colon in 1885 by German

bacteriologist Theodor Escherich. [2] Dr. Escherich also showed that certain strains of the

bacterium were responsible for infant diarrhea and gastroenteritis, an important public

health discovery. Although E. coli bacteria were initially called Bacterium coli, the name

was later changed to Escherichia coli to honor its discoverer.

E. coli is often referred to as the best or most-studied free-living organism. More

than 700 serotypes of E. coli have been identified. The “O” and “H” antigens on the
bacteria and their flagella distinguish the different serotypes. It is important to remember

that most kinds of E. coli bacteria do not cause disease in humans. Indeed, some E.

coli are beneficial, while some cause infections other than gastrointestinal infections,

such as urinary tract infections. (MarlerClarkAbout E. coli — Copyright © 2005-2017

Outbreak, Inc., All Rights Reserved)

Although most strains of E. coli bacteria are harmless and live in the intestines of

healthy humans and animals, several strains can produce powerful toxins and cause

severe illness in humans. This versatile pathogen is best known for being transmitted to

humans through contaminated foods — such as undercooked meat and unpasteurized

fruit juice — and has attracts much attention when serious outbreaks occur.

E. coli is capable of causing a wide variety of diseases — from urinary tract

infections to meningitis. A considerable amount of media coverage has recently been

devoted to one particular strain of E. coli, responsible for an estimated 73,000 cases of

infection and 61 deaths in the United States each year. Knowing more about the biology,

the evolution, and the genetic basis of this pathogen is crucial to future prevention of

infection and illness. (MichaelS.Donnenberg ISBN: 978-0-12-220751-8)

Related Studies

This study is related to the study entitled “Cytotoxic and Antibacterial Activity of

Basella Alba Whole Plant: A Relatively Unexplored Plant” performed by the researchers

RatheeSushila , AhujaDeepti , RatheePermender , ThankiMadhavi , RatheeDharmender

of PDM College of Pharmacy, Bahadurgarh, Haryana 2010

According to their study, The Cytotoxic activity was carried out using different

dilutions of methanolic extract against Jurkat and lung cancer cell lines. The antibacterial
activity was carried out using different dilutions of methanolic extract against gram

positive strains (Staphylococcusaureus,Micrococcusluteus,Bacillussubtilus) and gram

negative ones (Pseudomonasaeruginosa) by the cup-plate assay method and minimum

inhibitory concentrations (MICs). The TLC fingerprinting reveals the presence of β-

Sitosterol&Lupeol in the methanolic extract. The BAE showed significant dose

dependant cytotoxicity on Jurkat cell lines as compared with the reference standard. The

different concentrations of extract showed moderate activity against

Pseudomonasaeruginosa,Bacillussubtilis while weak response against

Staphylococcusaureus,Micrococcusluteus&Escherichiacoli. The minimum inhibitory

concentration of methanolic extract was 6.25µg/ml against

Staphylococcusaureus,Micrococcusluteus,Pseudomonasaeruginosa&Bacillussubtilus and

12.5µg/ml against Escherichiacoli. The methanolic extract shows the significant growth

inhibition on human cancer cell lines & momentous zone of inhibition for

microorganisms studied. The overall result of this study indicates that the methanolic

extract from Basella alba have interesting anticancer & antibacterial properties and the

traditional use of this plant may also derive from its antibacterial & anticancer properties.

This study is related to the study entitled “An Overview on Phytochemical, Anti-

Inflammatory and Anti-Bacterial Activity of Basella alba Leaves Extract” of A.K. Azad,

W.S. Wan Azizi, Z.M. Babar, Zubair Khalid Labu and S. Zabin 2013.

According to the study, Inflammation is the complex biological response of

vascular tissue to harmful stimuli such as pathogens, damaged cells, or irritants. In the

present investigation an attempt was made to screen the anti-inflammatory activity of

Basella alba leaf. From this study, it can be concluded that, Basella alba possesses a
good anti-inflammatory activity. In the cotton pellet induced inflammation and in the

carrageenan Induced inflammation the animals treated with the plant extract have been

shown a significant activity at 500 mg/kg dose (p<0.001) which was comparable with the

standard drug. In view of the percentage inhibition also the plant was studied, in which

the plant extract was found effective. The antibacterial activity was carried out using

different dilutions of methanolic extract against gram positive strains (Staphylococcus

aureus Micrococcus luteus, Bacillus subtilus) and gram negative ones (Pseudomonas

aeruginosa) by the cup-plate assay method and minimum inhibitory concentrations

(MICs). The different concentrations of extract showed moderate activity against

Pseudomonas aeruginosa, Bacillus subtiliswhile weak response against Staphylococcus

aureus, Micrococcus luteus and Escherichia coli. The minimum inhibitory concentration

of methanolic extract was 6.25µg/ml against Staphylococcus aureus, Micrococcus luteus,

Pseudomonas aeruginosa and Bacillussubtilus and 12.5 µg/ml against Escherichia coli.

The overall result of this study indicates that the methanolic extract of Basella alba have

interesting anti-inflammatory and antibacterial properties.

 CHAPTER III

METHODOLOGY

This chapter includes the information concerning the research design and the

materials used in conducting the research study. The methods, procedure and the

variables used in this study.

Research Design

The study will make use of experimental research method using one way

ANOVA (Analysis of Variance). The experimental method involves manipulating one

variable to determine if changes in one variable cause changes in another variable. This

procedure will be further studied using phytochemical screening to determine the active

properties present in the plant.

The ethanolic extract of Malabar Nightshade leaf was tested to determine the

susceptibility against the selected gram negative bacteria, antibacterial susceptibility

which make use for testing of pure culture of E.coli in the experiment to measure the

zone of inhibition of 100%, 75%, 50%, and 25% a paper disc soaked with the Malabar

Nightshade leaf ethanolic extract and will be compare to Ciprofloxacin as the positive

control.

The study is composed of three phases. Phase I was the Phytochemical Screening

analysis, Phase II was the Extraction Method, Phase III was the Disc Diffusion Method

and Phase IV was the treatment preparation.

Phytochemical Screening analysis

The phytochemical screening is conducted at Lyceum Northwestern University.

The result indicated the presence of alkaloidal salt, saponin, glycosides and non-

hydrolizable tannins. It shows that Malabar Nightshade leaves has an Antibacterial

Property against E. coli.

Preparation of Malabar Nightshade leaf ethanolic Extract

The Malabar Nightshade leaves were collected and washed Malabar Nightshade
leaves was cut into small pieces and weigh at 100 grams on the weighing scale. Then, it
was placed into an Erlenmeyer flask with 300 ml of ethanol for Maceration. Boil it for 1
hour and then filtrate it to condair the liquid content of the extract. Afterwards, place it to
evaporating dish for evaporation to get the crude extract.

Treatment Preparation

The following treatment concentrations were prepared:

Treatment 0 (T0) – Positive (+) control, Ciprofloxacin

Treatment 1 (T1) – 100% concentration of Malabar Nightshade leaf ethanolic extract

Treatment 2 (T2) – 75% concentration of Malabar Nightshade leaf ethanolic extract

Treatment 3 (T3) – 50% concentration of Malabar Nightshade leaf ethanolic extract

Treatment 4 (T4) – 25% concentration of Malabar Nightshade leaf ethanolic extract

Preparation of culture media

The nutrient agar for bacterial slants was prepared by dissolving a specific amount

of Mueller Hinton Agar in exact amount of distilled water. Mueller Hinton Agar was
used as the base agar for the bioassay. The standard Mueller Hinton Agar medium was

prepared by dissolving Mueller Hinton Agar in distilled water.

Disc Diffusion Method

In the preparation of inculation, pick around 3-5 well isolated colonies of the E.

coli to use for the brain heart infusion broth. Then incubate the E. coli at 37 ˚C for 2-8

hours until light to moderate growth of tarbidity appears. Inoculate the Mueller hinton

agar plate by using sterile cotton swab immersed into the inoculum (squeezing the swab

against the tube wall to remove the excess) by streaking. By using sterile forcep pick the

paper disc soaked with the Malabar Nightshade leaf ethanolic extract and distribute it to

the inoculated agar by pressing them into firm contact. Incubate the plate with E. coli at

37 ˚C for 24 hours. After 24 hours of incubation, observe the zone of inhibition

measuring it by Bernier caliper. Lastly, Record the result.

Bacterial Disposal

Add 250 milliliters of water to orange Biohazard Bag before closing. This is to

ensure proper sterilization. Use proportionately less water for smaller bags. Place in pan

suitable for transporting and autoclaving. Process for 60 minutes if using a metal pan and

75 minutes if using an autoclavable Nalgene pan at 121°C (15 psi). 4. Cool, then discard

autoclaved bag as general waste in the heavy clear plastic bags in the bins provided in the

autoclave area. Housekeeping will remove the autoclaved bags ONLY if they have been

properly autoclaved and placed into the clear bags. They will not remove any bags left on

the floor or in the autoclave.

Gathering of Data

Plant Collection: The leaves of Malabar Nightshade (Basella alba) are gathered from

Guesang, Mangaldan, Pangasinan and identified and certified by municipal agricultural

office of Mapandan, Pangasinan.

Bacteria: The experiment will use gram-negative bacteria which is the E.coli. These will

be cultured and obtained from Virgin Milagrosa University Foundation, San Carlos City,

Pangasinan.

Control and Reagents: Ciprofloxacin, a group of fluoroquinolones antibiotics drug will

serve as the positive control. This will be purchased from Virgin Milagrosa University

Foundation, San Carlos City, Pangasinan. The negative control is the Malabar Nightshade

leaf ethanolic extract used in the experiment.

Statistical Analysis

The data was summarized using ANOVA in and further analyzed to find out its

significant difference among the four different treatments with four different

concentrations used to test the effectiveness of the antibacterial property of Malabar

Nightshade leaves extract against E. coli

 FLOWCHART OF THE STUDY

Gathering of materials

Preparation of Malabar Nightshade leaves

Treatment Preparation

Disc Diffusion Method

Drawing Conclusions

Making Recommendations
 CHAPTER IV

RESULTS AND DISCUSSION

This chapter presents the result and findings of this study including its

corresponding analysis and interpretations.

Table 1: Phytochemical Screening result

Table 1.1 ALKALOIDS

Preliminary Test Result

Mayer’s regeant Yellow solution with brown precipitate
Wagner’s regeant Brown solution with brown precipitate

Confimatory Test
Mayer’s regeant Light yellow solution
Wagner’s regeant Yellow solution

Quaternary
Mayer’s regeant Yellow solution with brown precipitate
Wagner’s regeant Yellow solution with brown precipitate

Table 1.1 indicates the presence of alkaloidal salt due to the formation of

precipitate in the Preliminary and Quaternary tests.

Table 1.2 SAPONIN GLYCOSIDES

Result
Froth test Honeycomb formation
Liebermann-Burchard Test Reddish brown solution

Table 1.2 indicates the presence of saponin glycosides due to the formation of

honeycomb in Froth and color change in the Liebermann-Burchard test.

Table 1.3 TANNINS

Result
Gelatin test Yellow solution with precipitate
Ferric Chloride Test Greenish brown solution

Table 1.3 indicates the presence of non-hydrolyzable tannins due to the formation

of precipitate in the Gelatin test and greenish brown solutin in the Ferric Chloride test.

Table 2: Zone inhibition of E. coli in four different treatments and the positive

control.

Zone inhibition of E. coli in four different treatments and the positive control
Zone of inhibition of E. coli
SET UP
Trial 1 Trial 2 Trial 3
Treatment 1
(100 % Malabar Nightshade leaf 16 mm 16 mm 14 mm
ethanolic extract)
Treatment 2
(75 % Malabar Nightshade leaf 16 mm 16 mm 14 mm
ethanolic extract)
Treatment 3
(50 % Malabar Nightshade leaf 14 mm 14 mm 16 mm
ethanolic extract)
 Treatment 4
(25 % Malabar Nightshade leaf 14 mm 14 mm 16 mm
ethanolic extract)
Treatment 0
22 mm 22 mm 22 mm
(Positive control: Ciprofloxacin)

Table 2 shows the zone of inhibition of E. coli in four different treatments and the

positive control. It shows that treatment 0 has the highest zone inhibition of E. coli of 22

mm in three trials. While treatments 1 and 2 both have 16 mm in trial 1 and trial 2, and 14

mm in trial 3. And treatments 3 and 4 both have 14 mm in trial 1 and trial 2, and 16 mm

in trial 3.

Table 3: Mean average of zone of inhibition of E. coli in four different treatments

and the positive control.

Treatment Mean

Treatment 1 – 100% Malabar nightshade leaf ethanolic extract 15.33 mm

Treatment 2 – 75% Malabar nightshade leaf ethanolic extract 15.33 mm

Treatment 3 – 50% Malabar nightshade leaf ethanolic extract 14.66 mm

Treatment 4 – 25% Malabar nightshade leaf ethanolic extract 14.66 mm

Treatment 0 – Ciprofloxacin 22 mm

Table 3 shows the mean average of zone of inhibition of E. coli in four different

treatments and the positive control. It shows that treatment 0 which is the positive control

obtained the highest mean average of 22, while treatments 1 and 2 both have 15.33 mean

average and treatments 3 with treatment 4 have 14.66 mean average.

Table 4: Mean percentage of zone of inhibition of E. coli in four different treatments

and the positive control.

Treatment Mean Percentage

Treatment 1 – 100% Malabar Nightshade leaf ethanolic

15.33 80%
extract
Treatment 2 – 75% Malabar Nightshade leaf ethanolic
15.33 80%
extract
Treatment 3 – 50% Malabar Nightshade leaf ethanolic
14.66 75%
extract
Treatment 4 – 25% Malabar Nightshade leaf ethanolic
14.66 75%
extract

Treatment 0 – Ciprofloxacin 22 95%

Table 4 shows the mean percentage of zone of inhibition of E. coli in four

different treatments and the positive control. It can be gleaned from the table that

treatment 0 which is the positive control has 95% susceptibility. While treatments 1 and 2

both have 80% susceptibility. And treatments 3 and 4 both have 75% susceptibility.

Table 5: One-way ANOVA (Analysis of Variance)

One-way ANOVA (Analysis of Variance) TABLE

Sources of Sum of Mean
df F computed F tab Sig
Variation Squares Squares
Between
4 118.92 29.73 27.84 3.48 0.05
Group
Within
10 10.68 1.068
Group
TOTAL 14 129.6
 Table 5 shows the One-way ANOVA (Analysis of Variance). It can be gleaned

from the table that the Null Hypothesis (H0) is rejected Since the F ratio value of 27.84 is

greater than the F-tabular value of 3.48 at 0.05 level of significance. This reveals that

there is a significant difference between the four different treatments and the positive

control.
 CHAPTER V

SUMMARY, CONCLUSIONS, AND RECOMMENDATIONS

This chapter summarized the findings, presents the conclusions that was drawn

from the results and findings and the recommendation that is derived from the

conclusions.

Summary

This research study is entitled “Antibacterial Activity of Malabar Nightshade

(Basella alba) Leaf Ethanolic Extract against E. coli (Escherichia coli)” focused on the

study of the susceptibility of Malabar Nightshade leaf ethanolic extract in E. coli

(Escherichia coli). Phytochemical screening result indicates the presence of alkaloidal

salt, saponin glycosides and non-hydrolizable tannins enabling it to have antibacterial

property. Both treatment 1 and treatment 2 with 80% susceptibility of its antibacterial

property yields best. There is a significant difference between the antibacterial property

of Malabar Nightshade leaf ethanolic extract and the positive control which is the

Ciprofloxacin.

Conclusions

Based from the results accomplished throughout the study, the following

conclusions are formulated:

 1. Alkaloidal salt, saponin glycosides and non-hydrolizable tannins are the

significant active constituents present in the Malabar Nightshade leaf ethanolic

extract which certifies that it has antibacterial property.

2. The 100% and 75% pure concentration of Malabar Nightshade leaf ethanolic

extract with 80% susceptibility of its antibacterial property yields best among the

treatment used.

3. There is a significant difference between the antibacterial property of Malabar

Nightshade leaf ethanolic extract and the positive control. Since the F test values

of the four treatments versus the positive control (T1 vs T0, T2 vs T0, T3 vs T0, T4

vs T0) are greater than the critical value of 111.36, the significant difference is

between all the treatments and the positive control.

Recommendations

Based on the results of the study, the used of Malabar Nightshade leaves are

highly recommended to a person suffering from disease or infection caused by E. coli

bacteria. It is also economical and safer than the commercial anti-bacteria found in the

market.

The researchers recommend the cultivation of this plant from a prevalent study of

its other parts that should be commenced to find out the other properties that can be used

for prevention of common diseases and infections.

 BIBLIOGRAPHY

PRIMARY SOURCE

1. Middle-East Journal of Scientific Research, 14 (5). pp. 650-655. ISSN 1990-9233

2. RatheeSushila , AhujaDeepti , RatheePermender , ThankiMadhavi ,

RatheeDharmender of PDM College of Pharmacy, Bahadurgarh, Haryana 2010

3. A.K. Azad, W.S. Wan Azizi, Z.M. Babar, Zubair Khalid Labu and S. Zabin 2013.

SECONDARY SOURCES

1. Plant use (M. Ramansayah)

2. International Journal of Pharmacy and Pharmaceutical Research

3. About E. coli (Marler Clark)

Consultants

1. Marivic Paris, Microbiology Department of VirgenMilagrosa University

Foundation

2. Elizabeth Santonia, College of Pharmacy of VirgenMilagrosa University

Foundation

3. Alvin Franza, College of Pharmacy of VirgenMilagrosa University Foundation

4. Cesar Ramirez, Science Department of Mapandan National High School

 APENDICES

Appendix A. Gantt chart

Duration July-March 2017

Activities
Jun. Jul. Aug. Sept. Oct. Nov. Dec. Jan. Feb.
Identifying the
problem.
Finding the
solutions.
Searching for
the RRL
Planning the
experiment.
Setting the
experimental
design.
Purchase of
material.
Preparing the
materials
needed.
Gathering and
recording.
Analysis of Data
Writing Process