Chapter 4

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.

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.

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.

Objective 2. Determine the phytochemical properties of Manila palm

(Adonidia merrillii) fruit.

Table 2. Phytochemical Properties of Manila palm (Adonidia merrillii) Fruit.

Phytochemicals Flavonoid Tannins Saponins

Ethanolic crude extract of Manila palm ✓ ✓ ✓

fruit
 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 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 75% concentration and  100% concentration.

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 3.1. 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

EXPERIMENTA REP 1 REP 2 REP 3 MEAN INTERPRETATIO

L GROUP N

100% 0mm 0mm 0mm 0mm Resistant

Concentration

75% Concentration 0mm 0mm 0mm 0mm Resistant

CONTROL

GROUP

Positive control 31mm 29mm 29mm 29.7m Suceptible

(Ciprofloxacin) m

Negative control 0mm 0mm 0mm 0mm Resistant

(Distilled Water)
 STAPHYLOCOCCUS AUREUS

EXPERIMENTA REP 1 REP 2 REP 3 MEAN INTERPRETATIO

L GROUP N

100% 14mm 16mm 15mm 15mm Suceptible

Concentration

75% Concentration 13mm 17mm 17mm 15.67 Suceptible

mm

CONTROL

GROUP

Positive control 28mm 28mm 28mm 28mm Suceptible

(Ciprofloxacin)

Negative control 0mm 0mm 0mm 0mm Resistant

(Distilled Water)

Table 3.1. 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 and

Staphylococcus aureus. 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 (0 millimeter), indicating that the

replicates were resistant to the ethanolic crude extract. Only the positive control

(Ciprofloxacin) showed a powerful average of inhibition of 29.3 mm, indicating that

the replicates were susceptible to crude extract. On the microorganism,

Staphylococcus aureus, the three replicates using 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 (0 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 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 anti-boutiques, antimicrobial chemical compounds, and

even many herbal drugs. Gram-negative bacteria's relative resistance can be attributed

to their lipopolysaccharide layer and periplasmic space.

Table 3.2. Test Statistics of the positive control group and the experimental

group against Staphylococcus aureus.

Test Statisticsa,b

CIPROF EX100 EX75

Kruskal-Wallis H .000 2.000 2.000

df 2 2 2

Asymp. Sig. 1.000 .368 .368

a. Kruskal Wallis Test

b. Grouping Variable: REPLICATE

Table 3.2. 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.

STAPHYLOCOCCUS AUREUS

EXPERIMENTAL REP 1 REP 2 REP 3 MEAN INTERPRETATION

GROUP

100% Concentration 14mm 16mm 15mm 15mm Suceptible

75% Concentration 13mm 17mm 17mm 15.67 Suceptible

mm

ESCHERICHIA COLI

EXPERIMENTAL REP 1 REP 2 REP 3 MEAN INTERPRETATION

GROUP

100% Concentration 0mm 0mm 0mm 0mm Resistant

75% Concentration 0mm 0mm 0mm 0mm Resistant

Objective 4. Compare the efficacy of the antibacterial activity of Manila

palm fruit extract in terms of 75% concentration and 100% concentration. 

 Table 4.1. Efficacy of the antibacterial activity of Manila palm fruit

extract in terms of 75% concentration and 100% concentration against

Escherichia coli and Staphylococcus aureus.

Table 4.1 shows the efficacy of the antibacterial activity of Manila palm fruit

extract in terms of 75% concentration and 100% concentration against Escherichia

coli and Staphylococcus aureus. 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 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.

Given the findings, only Staphylococcus aureus exhibited antimicrobial

activity, being susceptible to the two concentrations (75% and 100%) of the Manila

palm (Adonidia merrillii) fruit's ethanolic crude extract. 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%.

 Table 4.2. P-values of 100% and 75%

Concentration P-value

100% .368

75% .358

Table 4.2. showed the p-values of 100% and 75%, which are both .368. This

indicates that both concentrations have no significant difference. Which accepts

Hypothesis 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.

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. Hypothesis Test Summary of significant difference between the

positive control group, the experimental group, and the negative control group.
 Hypothesis Test Summary

Null Hypothesis Test Sig. Decision

1 The distribution of CIPROF Independent- 1.00 Retain the

is the same across categories Samples Kruskal- 0 null

of Replicate Wallis Test hypothesis

2 The distribution of EX100 is Independent- .368 Retain the

the same across categories of Samples Kruskal- null

Replicate Wallis Test hypothesis

3 The distribution of EX75 is Independent- .368 Retain the

the same across categories of Samples Kruskal- null

Replicate Wallis Test hypothesis

4 The distribution of Independent- 1.00 Retain the

DISTILLED WATER is the Samples Kruskal- 0 null

same across categories of Wallis Test hypothesis

Replicate

Asymptotic significances are displayed. The significance level is .050.

Table 5. shows the significance of experimental and control groups. Both of

the experimental groups, which are the 100% and 75% concentrations, showed a

significance of .368. While both of the control groups, which are the positive group,

or Ciprofloxacin, and the negative group, or distilled water, showed a significance of

1.000.

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 Hypothesis 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.

Chapter 5

SUMMARY, CONCLUSION, AND RECOMMENDATIONS

This chapter presents the summary, conclusion, and recommendations of the

study.

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 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 and Staphylococcus aureus was obtained from Cotabato

Regional Medical Center.

To achieve the objectives of the study, the researchers assigned the

microorganisms of interest into 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 when it was subjected to the formulation were all the same for the two

microorganisms. This is to determine which of them 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)

was 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 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 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

susceptible level. The two concentrations from all the replicates of Escherichia coli,

on the other hand, showed no antibacterial inhibition with 0 millimeter, 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

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. Since.368 is above 0.50, 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 exhibited antimicrobial activity as being susceptible to the ethanolic crude

extract. It did not exhibit antimicrobial efficacy against Escherichia coli. 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. 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. Add more trials (up to three trials) and concentrations (such as 25% and 50%) of

the Manila palm (Adonidia merrillii) fruit’s ethanolic crude extract.