ANTIBACTERIAL ACTIVITY OF MANILA PALM (Adonidia merrillii (Becc.) Becc.

)
FRUIT EXTRACT AGAINST Escherichia coli AND Staphylococcus aureus

MARIA BJ THERESA A. AQUE

ALAN LOUISE Y. BUALAT
UMAYNAH E. AHMED

JULY 2023
ANTIBACTERIAL ACTIVITY OF MANILA PALM (Adonidia merrillii (Becc.) Becc.)
FRUIT EXTRACT AGAINST Escherichia coli AND Staphylococcus aureus

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A Thesis Presented to the

Faculty of the College of Medical Laboratory Sciences
Liceo de Cagayan University
Cagayan de Oro City

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In partial fulfillment of the

Requirements for the Degree
Bachelor of Science in Medical Laboratory Science

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MARIA BJ THERESA A. AQUE

ALAN LOUISE Y. BUALAT
UMAYNAH E. AHMED

JULY 2023
ANTIBACTERIAL ACTIVITY OF MANILA PALM (Adonidia merrillii (Becc.) Becc.)
FRUIT EXTRACT AGAINST Escherichia coli AND Staphylococcus aureus

By:

Maria BJ Theresa A. Aque

20200117546
maque17546@liceo.edu.ph

Alan Louise Y. Bualat

20200081913
abualat81913@liceo.edu.ph

Umaynah E. Ahmed
20200101568
uahmed01568@liceo.edu.ph

Adviser:

Marilou O. Honculada, RMT, MA, MSMLS

LICEO DE CAGAYAN UNIVERSITY

Cagayan de Oro City

Gunning Fog index: Original:

Flesch Reading Ease: Plagiarisms:
 ABSTRACT

The use of plants as medicine for fighting infections is controversial. The purpose of this
study was to determine the effectiveness of Manila palm (Adonidia merrillii (Becc.) Becc.)
ethanolic fruit extract in the inhibition of growth of Escherichia coli and Staphylococcus aureus.
The ethanolic fruit extract was obtained through rotary evaporation and was screened for
phytochemical compounds using qualitative methods. Two concentrations of the fruit extract were
then impregnated to filter paper discs, namely 75% and 100% concentration, with other discs
containing ciprofloxacin as the positive control and distilled water as the negative control. The
bacterial colonies of E. coli and S. aureus were streaked in Mueller Hinton Agar plates for
susceptibility testing in hand with the experimental and control groups and to compare the zone of
inhibition of the fruit extract against E. coli and S. aureus. Testing was done in triplication to ensure
the efficacy of the fruit extract. The extract exhibited a presence of flavonoids, tannins, and
saponins. The fruit extract exhibited a zone of inhibition against S. aureus with a mean of 15mm
and 15.67mm for 100% and 75% concentration respectively which is above 10mm; indicating that
the bacteria was susceptible against the ethanolic fruit extract of Manila palm.

Keywords: Escherichia coli, ethanolic extract, flavonoids, Manila palm, phytochemical screening,
Staphylococcus aureus, susceptible, zone of inhibition

THE PROBLEM AND ITS SCOPE

Introduction

Plants remain imperative in the continuous survival of human beings. It fulfills our basic
needs for living, especially in the present day where environmental and health problems seem to be
the “new normal”. With the diverse purposes that plants have, one of them is being a medicinal and
therapeutic alternative. Herbal medicine exists globally, with some cultures even tracing its
medicinal use for thousands of years. Although pharmaceutical drugs are now available in the
market, numerous people still choose to use herbal medicine as their primary plan of action for
treatment. Moreover, people are recently leaning more into organic products and effusing herbal
life into their daily living, making it a huge trend these days. Despite plants' beneficial attributes,
there is still plenty of room in the research community that will support plants’ effectiveness
towards fighting infection, specifically the utilization of the fruit of Manila Palm.
Over the years, clinical and technological advancements have given us an array of options
on how we can deal with bacterial infections, such as antibiotic drugs. However, antimicrobial
resistance (AMR) incidences are having a continuous rise as microorganisms (such as bacteria,
fungi, viruses, and parasites) are responding to the antimicrobial drugs and developing AMR,
making pills lose their effectiveness (Zhu, Huang, and Yang, 2022). Backed up by the study of Rai,
Randhawa, and Kaur (2013), the global clinical problem might lead to the return of the pre-
antibiotic era, or in other words, herbal or plant-based medicine. 
Given the aforementioned facts, this research will significantly serve as an additional
reference for the research community about the potential of this plant’s fruit extract and also
encourage them to use the other parts of the Manila palm for other beneficial purposes. This study
will also significantly contribute to the pharmaceutical section and may develop drugs
incorporating this plant’s fruit extract. Very few studies are available on Manila palm, which is why
delving more into its capabilities as a potential medicine against S. aureus and E. coli infections is
the purpose of this study, to battle the worrying resistance of microorganisms with the available
medicinal drugs available today.
Theoretical and Conceptual Framework

This study was anchored with the study of Essein, et.al. (2017), where the Manila palm (Adonidia
merrillii) and Archontophoenix tukeri methanolic fruit extract was utilized and screened for
phytochemical compounds and its in vitro cytotoxic, antioxidant, and antimicrobial activity. The
extract showed a promising result where it exhibited antioxidant activity and cytotoxicity. In the
phytochemical screening, it was found out that flavonoids, phenols, and deoxysugars were highly
present in the methanolic fruit extract. According to Xie, Yang, Tang, Chen and Ren (2015),
flavonoids are well known for their antibacterial effect on many of pathogenic microorganisms by
inhibiting nucleic acid synthesis, cytoplasmic membrane function, and more.

Schematic Presentation

The schematic presentation presents the independent and dependent variables. The Independent
variables are composed of an experimental group which consists of 75% concentration and 100%
concentration and a control group which consists of positive control and negative control. The
Dependent variable is the antimicrobial activity of Manila palm.
.

Antimicrobial activity of
Manila Palm

Figure 1. The schema of the study that shows the interplay of all variable
Objectives of the Study

The study aims to determine the antibacterial activity of Manila palm (Adonidia merrillii)
fruit extract against E. coli and S. aureus.
Specifically, it aims to:
1. Collect, identify, classify, and describe the Manila palm (Adonidia merrillii) plant.
2. Determine the phytochemical property of Manila palm (Adonidia merrillii) fruit.
3. Determine the antibacterial activity of Manila palm (Adonidia merrillii) fruit extract using
the following:
1. 75% concentration
2.  100% concentration
3. Positive control (Ciprofloxacin)
4. Negative Control (Distilled water)
4. Determine the significant difference in the antibacterial activity of Manila palm fruit extract
in terms of:
1. 75% concentration; and 
2. 100% concentration. 
5. Determine the significant difference in the antibacterial activity of Manila palm (Adonidia
merrillii) fruit extract against the experimental and the control groups.

Null Hypothesis
Ho1: There is no significant difference in the antibacterial activity of Manila palm (Adonidia
merrillii) fruit extract with the different concentration:
1. 75% concentration
2. 100% concentration
Ho2: There is no significant difference in the antibacterial activity of Manila palm (Adonidia
merrillii) fruit extract against the experimental and the control groups.

Significance of the Study

The findings of this study might be expected to be essential and benefit the following
sectors:
Pharmaceutical. The study is expected to give pharmaceuticals the idea that there is a
natural herbal extract which provides an alternative antibacterial control which can fight diseases
caused by Escherichia coli and Staphylococcus aureus.
Community. The study is expected to provide awareness to people about the antibacterial
activity of Manila palm (Adonidia merrillii) fruit extract against Escherichia coli (bacteria found in
the intestines of healthy people and animals that can manifest clinically as abdominal cramps,
bloody diarrhea, and even kidney failure) and Staphylococcus aureus (bacteria found in the nose
and on the skin of humans, which is the most common cause of skin and soft tissue infections
including abscesses, furuncles, and cellulitis), allowing the community to consider organic
antibacterial controls.
Patients. The findings of study will help previously or currently infected patients with
Escherichia coli and Staphylococcus aureus to use plant-based antibacterial controls instead of
chemically synthesized ones.
Medical Laboratory Science Students. The study is expected to broaden the skills of
Medical Laboratory Scientists as well as researchers in terms of finding, analyzing, considering
different methods, and understanding the susceptibility of Escherichia coli and Staphylococcus
aureus to the antimicrobial activity of Manila palm (Adonidia merrillii) fruit extract.
 Future Researchers. Indeed, the progress and improvements of this study will be possible
with the help of the future researchers especially in developing another study that will utilize the
other parts of the Manila palm (Adonidia merrillii) for potential medicinal use.

Scope and Delimitations of the Study

This study was confined to the determination of the antibacterial activity of Manila Palm
(Adonidia merrillii) fruit extract against Escherichia coli and Staphylococcus aureus using two
concentrations: 75% and 100%. The primary part that was utilized in this study is the fruit of the
Manila palm and was collected in Barangay Victoria, Oroquieta City, Misamis Occidental,
Philippines. The actual testing of this research takes place at MLS Laboratory 2 & 3 of the College
of Medical Laboratory Science located on the Fourth Floor of Liceo de Cagayan University, Paseo
del Rio Campus, Rodelsa Circle, Macasandig, Cagayan de Oro City, Philippines. A timeline the
first and second semesters of the academic year 2022-2023 were set to cover the processes involved
in this research.

Definition of Terms
The following terms are operationally defined for a better understanding of this study.
Antimicrobial activity – This refers to the effectivity of Manila Palm (Adonidia merrillii)
fruit extract to inhibit the growth of E. coli and S. aureus
Ciprofloxacin – This will be used as a positive control of this study.
Concentration – In this study, this will be differentiated into 75% and 100%. 
Distilled Water – This will serve as the negative control of this study.
Escherichia coli – For this study, these bacteria will be experimented for the antibacterial
activity of Manila Palm (Adonidia merrillii) fruit extract. 
Ethanol – This will be used as the vehicle control of this study.
Negative Control – It is a solution that cannot inhibit the growth of E. coli and S. aureus.
Positive Control – It is a known antibiotic that can inhibit the growth of E. coli and S.
aureus.
Staphylococcus aureus – In this study, this will be the bacteria to be used for experimenting
the antibacterial activity of the fruit extract of Manila Palm (Adonidia merrillii).

MATERIALS AND METHODS

Research Setting
The Manila palm fruit (Adonidia merrillii) was collected at Barangay Victoria, Oroquieta
City in the province of Misamis Occidental. The experiment took place at MLS Laboratory 2 & 3
of the College of Medical Laboratory Science located at the Fourth Floor of Liceo de Cagayan
University, Paseo del Rio Campus, Rodelsa Circle, Macasandig, Cagayan de Oro City, Philippines.
The procedures were performed by the Medical Laboratory Science researchers and guided by a
registered medical technologist. The experiment used a pure culture of identified Escherichia coli
ATCC 25922 and Staphylococcus aureus ATCC 25923. The microorganisms were obtained from
the Cotabato Regional Medical Center. The Pharmaceutical Laboratory of Liceo de Cagayan
University was utilized to perform rotary evaporation and soxhlet with the company of registered
Pharmacist. 

Research Design
The study employed an experimental research design for it to utilize controls to compare the
efficacy of the antibacterial activity of Manila palm (Adonidia merrillii) fruit extract regarding
different concentrations against Escherichia coli and Staphylococcus aureus. 
 Collection and Sampling Procedure
In obtaining the bacterial sample, a pure culture of E. coli and S. aureus was taken from
Cotabato Regional Medical Cemter, Cotabato, Philippines, and put into sterilized tubes as its
primary container, and enclosed in an ice box as its secondary container.
The Manila palm fruit (Adonidia merrillii) fruit were collected within Brgy Victoria,
specifically at Oroqueita City, Philippines

Research Instrument
The researchers utilized the following materials and equipment during the experiment:
Manila palm fruit (Adonidia merrillii), pure culture of Escherichia coli and Staphylococcus aureus,
mortar and pestle, refrigerator for the bacterial culture and preservation, laboratory oven and
ethanol for the extraction of Manila palm fruit, rotary evaporator (rotavap), soxhlet, petri plates,
inoculating loops, alcohol lamps, electronic single pan balance, electric stove, incubator, autoclave,
autoclave tape, Whatman filter paper, Mueller-Hinton agar, water bath, Bunsen burner, wire gauze,
evaporating dish, Erlenmayer/Florence flask, beakers, volumetric flask, stirring rods, pipettes, test
tubes and test tube racks, forceps, ethanol or isopropanol and sodium hypochlorite for the
disinfection of working areas, Personal Protective Equipment (PPE), gadgets: laptop for data input
and storage and camera for documentation. 

Data Gathering Procedure

The researchers sought approval from the adviser after careful assessment and review of the
manuscript for the thesis. The researchers started conducting the experiment starting from the
collection of the Manila palm fruit (Adonidia merrillii) at Barangay Victoria, Oroquieta City. After
sample collection, the sample was sent to Central Mindanao University for Taxonomy Certification.
The researchers proceed with the actual experiment. The Manila palm fruit was powdered and
soaked with ethanol for extraction. The extract was tested at the Department of Pharmacy Liceo de
Cagayan University for Phytochemical screening. Once the extraction was made, it was diluted into
different concentrations which are 75% and 100% Manila palm fruit extract. The Mueller-Hinton
Agar were used for the antimicrobial susceptibility testing. The researchers obtained the identified
Escherichia coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923) at Cotabato Regional
Medical Center. The researchers prepared the inoculum with Escherichia coli (ATCC 25922) and
Staphylococcus aureus (ATCC 25923). The suspension is compared with 0.5 McFarland Standard
when the turbidity matches, the suspension is ready for inoculation. The inoculum was suspended
in the Mueller-Hinton Agar using the Streaking method to evenly distribute the bacteria in the plate.
The discs with different concentrations of Manila Palm fruit extract, positive control, negative
control and vehicle control were impregnated in the plate following the proper placement to prevent
overlapping. The researchers incubated the plates for 24 hours. After incubation the researchers
measured the zone of inhibition using a ruler. The results were interpreted into Resistant (R),
Intermediate (I) and Susceptible (S). Once results are released the researchers proceed in discussion
and provide a conclusion to the experiment.

Statistical Treatment and Analysis

Different statistical tool was employed to answer each objective of the study:
The first objective utilized Descriptive statistics for the collection, identification,
classification and description of Manila palm (Adonidia merrillii) plant.
 The second objective utilized Descriptive statistics for the determination of phytochemical
properties of Manila palm (Adonidia merrillii) fruit.
The third objective utilized Mean and Standard Deviation for the determination of
antibacterial effects of Manila palm (Adonidia merrillii) fruit extract in terms of different
concentrations. 
The fourth objective utilized Wilcoxon-Mann-Whitney U Testing to compare the efficacy of
the antibacterial activity of Manila palm (Adonidia merrillii) fruit extract in terms of different
concentrations. 
The fifth objective utilized Kruskal Wallis H Test to determine the significant difference of
the antibacterial activity of Manila palm (Adonidia merrillii) fruit extract against the experimental
and control groups. 
Furthermore, the researchers also used non-parametric tests such as the Wilcoxon-Mann-
Whitney U testing along with the Kruskal-Wallis t-test. These tests are contributory in cases where
two variables are not normally distributed. A comparison between the effectiveness of two
independent groups in a clinical trial will be easily noticed and distinguished. This assessed whether
the values in one group were greater or less than those in the other group.

PRESENTATION, ANALYSIS, AND INTERPRETATION OF DATA

This chapter presents the results of the antibacterial activity of Manila palm (Adonidia
merrillii) fruit extract in different concentrations, the negative control (distilled water), and the
positive control (Ciprofloxacin) against Escherichia coli and Staphylococcus aureus. The
presentation of data follows the logical order of the objectives of the study.

Objective 1. Collect, identify, classify and describe the Manila palm (Adonidia merrillii) plant.

Table 1.
Classification of Manila palm (Adonidia merrillii) Plant

Kingdom Plantae
Phylum Magnoliophyta
Class Liliopsida
Subclass Arecidae
Order Arecales
Family Arecaceae
Genus Adonidia
Species Merrillii
Other name Manila palm
The fruit of Adonidia merrillii, which was used as a sample in this study, initially grows
green, then turns orange to red when ripe, and has a small, ovoid appearance. In the study of Essien
et al. (2017), they described Adonidia merrillii fruit as consisting of a thin epicarp, a dry, yellowish,
thin-fleshy mesocarp, and a thin, fragile endocarp. Adonidia merrillii fruit is used as a masticatory
when ripe but is an inferior substitute for betel nuts. Adonidia merrillii was collected in the vicinity
of Misamis Occidental, specifically in Barangay Victoria, Oroquieta City, and has undergone plant
authentication on May 24, 2023. The plant sample was identified and confirmed by Dr. Maria
Melanie M. Guiang, a Botanist and member of the Pool of Taxonomists at Central Mindanao
University's Museum. The Manila palm (Adonidia merrillii) plant is classified in Table 1.
Objective 2. Determine the phytochemical properties of Manila palm (Adonidia merrillii)
fruit.

Table 2
Phytochemical properties of Manila palm (Adonidia merrillii) Fruit
Phytochemicals Flavonoids Tannins Saponins
Ethanolic crude extract of Manila palm fruit + + +
Note. (+) = present; (-) = absent
Table 2. shows the phytochemical properties of Adonidia merrillii fruit. An array of
phytochemical screening tests and procedures were performed on the fruit's ethanolic crude extract
to detect the presence of common compounds known to have antibacterial properties. The result of
the phytochemical screening tests and procedures revealed the presence of flavonoids, saponins,
and tannins. Flavonoids, one of the most abundant phytochemicals in Adonidia merrillii, were
identified by Ullah et al. (2020) as phytochemical compounds found in many plants, fruits,
vegetables, and leaves that have potential applications in medicinal chemistry. Also, there is
mounting evidence that flavonoids, saponins, and tannins have antibacterial activity against both
gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria
(Miklasińska-Majdanik et al., 2018).

Objective 3. Determine the antibacterial activity of Manila palm (Adonidia merrillii)

fruit extract using the experimental groups which are 75% and 100% concentration and
control groups which are positive control (Ciprofloxacin) and negative control (Distilled
water).
According to Iyasele et al., (2022), the result is determined by the diameter of the zone of
inhibition; a zone of inhibition measuring <7mm is resistant, 7-9mm is intermediate and >10mm is
susceptible. The results were interpreted as resistant (R), intermediate (I), or susceptible (S).

Table 3
Comparison between the zones of inhibition of Adonidia merrilli extract in millimeters (mm) by the
positive control, negative control and the experimental control group against Escherichia coli and
Staphylococcus aureus

ESCHERICHIA COLI
EXPERIMENTAL REP 1 REP 2 REP 3 MEAN INTERPRETATION
GROUP
100% Concentration 6mm 6mm 6mm 6mm Resistant
75% Concentration 6mm 6mm 6mm 6mm Resistant
CONTROL GROUP
Positive control 31mm 29mm 29mm 29.7mm Susceptible
(Ciprofloxacin)
Negative control 6mm 6mm 6mm 6mm Resistant
(Distilled Water)
Note. <7mm=Resistant; 7-9mm=Intermediate; >10mm=Susceptible
STAPHYLOCOCCUS AUREUS
EXPERIMENTAL REP 1 REP 2 REP 3 MEAN INTERPRETATION
GROUP
100% Concentration 14mm 16mm 15mm 15mm Susceptible
 75% Concentration 13mm 17mm 17mm 15.67mm Susceptible
CONTROL GROUP

Positive control 28mm 28mm 28mm 28mm Susceptible

(Ciprofloxacin)

Negative control 6mm 6mm 6mm 6mm Resistant

(Distilled Water)

Table 3 shows the comparison between the zones of inhibition of the experimental group
using different concentrations (100% and 75%), a positive control group (Ciprofloxacin), and a
negative control group (distilled water) of the Manila palm (Adonidia merrillii) fruit’s ethanolic
crude extract against Escherichia coli (ATCC 25922) and Staphylococcus aureus (ATCC 25923).
On measuring the inhibitions of Escherichia coli, the three replicates utilizing the 75% and 100%
concentrations and the negative control (distilled water) showed no antibacterial inhibition, with
only 6mm diameter, indicating that the replicates were resistant to the ethanolic crude extract. Only
the positive control (Ciprofloxacin) showed a powerful average of inhibition of 29.7 mm, indicating
that the replicates were susceptible to crude extract. On the microorganism Staphylococcus aureus
(ATCC 25923), the three replicates that used a 75% concentration showed an average inhibition of
15.67 millimeters, indicating that all replicates were susceptible to the extract. The three replicates
using 100% concentration showed an average inhibition of 15 millimeters, indicating that the
replicates also exhibited antimicrobial efficacy up to susceptible level. The three replicates utilizing
the negative control (distilled water) showed no antibacterial inhibition (6 millimeter), which makes
it resistant to the crude extract, while the positive control showed an average inhibition of 28
millimeters, indicating that all replicates were susceptible to the extract.
From the results of the table, it can be implied that of the two microorganisms, only
Staphylococcus aureus  (ATCC 25923) exhibited antimicrobial activity as being susceptible to the
Manila palm (Adonidia merrillii) fruit’s ethanolic crude extract. In fact, in the study of
Koohsari, Ghaemi, Sheshpoli, Jahedi, and Zahiri (2015), it shows that gram-positive bacteria are
more sensitive to herbal extracts than gram-negative bacteria. This could be due to gram-negatives'
inherent tolerance as well as the nature and composition of herbs. According to studies, the cell
walls of gram-positive bacteria are more sensitive to many antimicrobial chemical compounds and
even many herbal drugs. Gram-negative bacteria's relative resistance can be attributed to their
lipopolysaccharide layer and periplasmic space. Between the two concentrations the 75% showed a
higher mean of 15.6mm whereas the the 100% showed a mean of 15mm having a difference of
6mm. According to the results obtained in the study of Paz, Contreras, Munguía, Aguilar,
Inungaray (2018), prove that a higher percentage yield in obtaining the extract is not an indicator of
increased antibacterial activity. In general, the three extracts had a 75% efficiency for S. aureus and
S. typhi, 50% for S. paratyphi and 100% for E. coli. In the three extracts obtained by different
methods the percentage activity was 100% and the Index of Bacterial Susceptibility (IBS) was 75%.
Furthermore, in the study of Swiner (2022), he explained that since 75% have more water compared
to 100% that do not have water at all, it has the advantage of allowing it to dissolve more slowly,
penetrate cells, and kill bacteria.

Objective 4. Determine the significant difference in the antibacterial activity of Manila palm
fruit extract in terms of 75% concentration and 100% concentration. 

Table 4
Concentration Significance level
 100% .368
75% .368
Significance level of the two concentrations from Kruskal Wallis H Test.
Table 4 shows the significant difference of 100% and 75%, which are both .368. This
indicates that both concentrations have no significant difference. Which accepts Ho 1, indicating that
there is no significant difference in the anitibacterial activity of Manila palm (Adonidia merrillii)
fruit extract with the different concentrations. According to the study of Ambon et al. (2019), using
three different concentrations: 50%, 75%, and 100%, found that there was no significant difference
among the concentrations. Table 4 shows the efficacy of the antibacterial activity of Manila palm
fruit extract in terms of 75% concentration and 100% concentration against Escherichia coli
(ATCC 25922) and Staphylococcus aureus (ATCC 25923). In the zone of inhibition of Escherichia
coli, the three replicates utilizing the 75% and 100% concentrations showed no antibacterial
inhibition (0 millimeter), indicating that the replicates were resistant to the ethanolic crude extract.
In contrast, when the zone of inhibition of Staphylococcus aureus (ATCC 25923) was measured,
the three replicates using a 75% concentration showed an average inhibition of 15.67 millimeters,
indicating that all replicates were susceptible to the extract. In addition, the three replicates using
100% concentration showed an average inhibition of 15 millimeters, indicating that the replicates
also exhibited antimicrobial efficacy up to the susceptible level.

Objective 5. Determine the significant difference in the antibacterial activity of Manila

palm (Adonidia merrillii) fruit extract against the experimental and the control groups.

Table 5
Kruskal Wallis H Test with the corresponding significance level of the experimental groups and
control groups and their interpretations
Asymp. Sig. Interpretation
75% concentration .368 Retain null hypothesis
Ciprofloxacin 1.000 Retain null hypothesis
100% concentration .368 Retain null hypothesis
Distilled water 1.000 Retain null hypothesis
Note. Significance level is 0.050.
Table 5 contains the Kruskal-Wallis H test result, and it showed that there was no
statistically significant difference in pain score between the two different concentrations, namely
100% and 75% concentrations, having both a .368 pain score. Moreover, Ciprofloxacin has a pain
score of 1.000. In the study of Ambon, Guerra, Macapagal, Noble, Param, and Sta. Rosa (2019),
using three different concentrations: 50%, 75%, and 100%, found that there was no significant
difference among the concentrations.
This implies that there is no significant difference between the experimental group and the
control group. As both groups have exceeded the 0.05 significance level. Which accepts Ho 2,
indicating that there is no significant difference in the antibacterial activity of Manila palm
(Adonidia merrillii)  fruit extract between the experimental and control groups.
According to Tenny and Abdelgawad (2022), if the p value is higher than the significance
level, the null hypothesis is not refuted, and the results are not statistically significant. If the p value
is lower than the significance level, the results are interpreted as refuting the null hypothesis and
reported as statistically significant.
SUMMARY, CONCLUSION, AND RECOMMENDATION

Summary
The main objective of this study was to determine the antibacterial activity of Manila
palm (Adonidia merrillii) fruit extract against Escherichia coli and Staphylococcus aureus.
Furthermore, it aimed to determine whether there is a significant difference in the zone of inhibition
in each experimental group using the following concentrations of the fruit's ethanolic crude extract:
100% and 75%.
The fruit of Manila palm was collected in an area within the vicinity of Misamis Occidental,
specifically Barangay Victoria, Oroquieta City. The test organism, Escherichia coli (ATCC 25922)
and Staphylococcus aureus (ATCC 25923) was obtained from Cotabato Regional Medical Center.
To achieve the study’s objectives, the researchers assigned the microorganisms of interest to
two experimental groups. Both bacteria were subjected to the ethanolic crude extract, and their
susceptibility was measured separately. The amount of crude extract, the number of trials and
replicates, and the time and day it was subjected to the formulation were all the same for the two
microorganisms. This is to determine which is more sensitive to the ethanolic crude extract. The
two bacteria were subcultured separately on nutrient agar medium, and their exposure to different
concentrations (100% and 75% ethanolic crude extract), the positive control (Ciprofloxacin), and
the negative control (distilled water) were monitored. The differences in their susceptibility,
measured by the diameter of inhibition in millimeters, were attributed to the antimicrobial property
of the said extract.
Based on the results, only Staphylococcus aureus (ATCC 25923) showed antibacterial
activity at any of the two concentrations of the crude extract. The zones of inhibition based on the
antimicrobial property of the ethanolic crude extract against Staphylococcus aureus (ATCC 25923)
using three replicates of 75% concentration revealed an average inhibition of 15.67 millimeters,
indicating that all replicates were susceptible to the extract. The three replicates using 100%
concentration showed an average inhibition of 15 millimeters, indicating that the replicates also
exhibited antimicrobial efficacy up to a susceptible level. The two concentrations from all the
replicates of Escherichia coli (ATCC 25922), on the other hand, showed no antibacterial inhibition
at 6 millimetres, making it resistant to the crude extract. Only the positive control shows
antibacterial activity in all of the replicates. The mean of the susceptible was then computed. In
100% and 75% concentrations, the statistical data utilizing the Kruskal-Wallis H test showed no
statistically significant difference in pain score between the two different concentrations, namely
100% and 75% concentrations having both a .368 significance level. Since .368 is above 0.050, the
null hypothesis is accepted. The significance of the control group, both positive and negative, was
1.000, which accepts the null hypothesis.

Conclusion
From the findings of the study, it can be inferred that from the two concentrations of the
crude extract, the 75% and 100% concentrations of Manila palm (Adonidia merrillii) fruit’s
ethanolic crude extract, only Staphylococcus aureus (ATCC 25923) exhibited antimicrobial activity
as being susceptible to the ethanolic crude extract. It did not exhibit antimicrobial efficacy against
Escherichia coli (ATCC 25922). Furthermore, both hypotheses are accepted. Hence, there is no
significant difference in the antibacterial activity of Manila palm (Adonidia merrillii) fruit extract
with the different concentrations of 75% and 100%. And there is no significant difference in the
antibacterial activity of Manila palm (Adonidia merrillii) fruit extract between the experimental and
control groups.

Recommendations
Based on the findings of the study, the following recommendations are suggested: 
1. Utilize other microorganisms in order to determine the antibacterial activity of Manila palm
(Adonidia merrillii) fruit’s ethanolic crude extract.
2. For the pharmaceutical industry, conduct further research: invest in scientific studies to
determine the efficacy, safety, and appropriate dosage of Manila palm fruit ethanolic extract
against different strains of Escherichia coli. To use the other parts of the Manila palm
(Adonidia merrillii), such as the bark, flowers, leaves, stem, and roots, to create an extract that
kills microorganisms other than bacteria.
3. For Community, examine, research, and assess the various components of the manila palm and
note its various features to gain a deeper understanding of the plant.
4. For Patients, look into the plant's potential as a treatment for various bacterial infections.
Review the literature on the efficacy of the plant's other parts.
5. For the Medical Laboratory Science Students, review available scientific literature, investigate
the possibility of combining Manila palm fruit ethanolic extract with other antimicrobial agents
to see if there are any synergistic effects, and emphasize good laboratory practices such as
proper handling, preparation, and storage of Manila palm fruit ethanolic extract.
6. For the future researchers, conduct a comprehensive literature review, consider exploring
different extraction techniques to determine the optimal method that yields consistent results,
and perform in vitro studies to assess the antimicrobial activity of Manila palm fruit ethanolic
extract against E. coli. and S. aureus, and consider adding more trials (up to three trials) and
concentrations (25% and 50%) of the Manila palm (Adonidia merrillii) fruit’s ethanolic crude
extract.

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