Antimicrobial Activity of Bird’s Nest Fern Extract Against Escherichia Coli (an Experimental Research)

Republic of the Philippines

Sultan Kudarat Polytechnic State College

Science Laboratory High School

ACCESS, EJC Montilla, Tacurong City

This study

Antimicrobial Activity of Bird’s Nest Fern Extract Against

Escherichia Coli

is presented to Division Science Fair 2009.

RESEARCHERS:

Breylle Bepitel

Earl Adam Prado

Rona Angelique Manuel

RESEARCHER ADVISER:

Prof. Merlyn Leysa

September 2009

Dedication

This humble work is dedicated to all

Who have committed themselves

To the improvement

Of the life of our people.

ACKNOWLEDGEMENT

The researchers recognize with the gratitude and appreciation their indebtness to the

following persons and institutions who have made valuable contribution to the successful

completion of this study:

To the Prof. Merlyn Leysa, our researcher adviser for her time, for the guidance,

patience in checking drafts, encouragement and for giving enough consideration;

To the SKPSC faculty and staff and Dr. Herminia Dumadag for the untiring support

extended to us;

To the SKPSC Research Center , who provided us the materials and equipment used in

the conduct of this study;

To our classmates and friends who provided from the companionship and inspiration for

the completion of this study;

To our beloved parents who are supporting us for helping and providing all we need

and;

Most of all, God, who has endowed the researchers with intellectual gifts and strong

faith which sustained them through the different stages of their research work.

The Researchers

ABSTRACT

Breylle Bepitel, Earl Adam Prado, and Rona Angelique Manuel. A project design

“Antimicrobial Activity of Bird’s Nest Fern Extract Against Escherichia Coli”. Sultan Kudarat

Polytechnic State College- Laboratory High School, Research Center , EJC Montilla, Tacurong

City . September 2009.

Advisers: Prof. Merlyn Leysa.

The study Antimicrobial Activity of Bird’s Nest Ferns Extract against Escherichia Coli”

generally aimed to determine the potential of bird’s nest fern as an antibiotic for Escherichia

Coli.

Specifically the study aimed to:

1. Determine the sensitivity of E. coli to bird’s nest fern extract through Kirby-

Bauer antibiotic testing.

2. Determine the treatment where the specimens of bacteria are most sensitive to

3. Determine the significant differences among treatments and replications

conducted.

The major findings of this study were as follows:

1. Treatment 1(pure birds nest extract) got a general mean of its ZI with 59.45 mm while

Treatment 2 (5 ml bird’s nest extract + 5ml of sterilized water) got a general mean of

0.67mm. The control cefuroxime got a general mean of 51.11mm.

2. Escherichia coli showed high sensitivity when exposed to pure bird’s nest fern extract.

3. It is therefore found out that the treatment 1 is where Escherichia coli is most

sensitive to.

4. Based on the Analysis of Variance, treatments have significant differences among

each other in all trials conducted, while the replicates and trials are not significant.

After the 24-hour incubation, the diameter of the zone of inhibition of E. coli were

determined as positive or negative to be sensitive to bird’s nest fern.

Based on the findings of the test conducted, bacteria E. coli is favorably sensitive to

treatment 1, which is composed of pure bird’s nest extract from its leaves as it were remarked

sensitive. The treatments showed highly significant differences while the replicates and trials

are very precise and showed not much significance. Likewise, the comparison between the

control group and the treatments used cannot be directly measured since the control

(cefuroxime) is manufactured considering the dosage for human consumption while bird’s nest

fern extract is made to combat E. coli in direct contact with the bacteria itself.

Nevertheless, the study generally proved the great potential of Bird’s Nest fern extract

to be the radical antidote for the threatening public health nightmares, the E. coli.

TABLE OF CONTENTS

TITLE PAGE ………………………………………………………………………….i

Dedication ……………………………………………………………………………..ii

Acknowledgement……………………………………………………………………..iii

Abstract …………………………………………………………………………….....iv

THE PROBLEM ……………………………………………………………………..1

Background of the Study ……………………………………………………………..1

Objectives …………………………………………………………………………....2

Significance of the Study ……………………………………………………………...3

Hypotheses …………………………………………………………………………...3

Scope and Delimitations ……………………………………………………………....3

Definition of Key terms ..……………………………………………………………...4

REVIEW OF RELATED LITERATURE AND STUDIES …………………...........5

RESEARCH METHODOLOGY…………………………………………………. ….12

Research Design …………………………………………………………………....12

Material Gathering Procedure…..………………………………………………............12

Preparation Of Experiment Set-Up….….………………………………………...……12

Materials and Equipment ….…………………………………………………………...13

Methodology .. ………………………………………………………………………..14

Data Collection Procedure …………………………………………………………..15

Statistical Analysis …………………………………………………………………......15

RESULTS AND DISCUSSIONS…….……………………………………………….16

SUMMARY OF FINDINGS, CONCLUSION, AND RECOMMENDATION………24

Findings ……………………………………………………………………………….24

Conclusions ……………………………………………………………………….......25

Recommendations……………………………………………………………………...25

APPENDICES

LITERATURE CITED

CHAPTER 1

The Problem

Modernization like advancement in many fields grow faster, definitely people are aware

of this. It leads to open their minds to discover and expand their knowledge through developing

their concepts to solve different health problems.

Today, antibiotic resistance is one of the major problems of medical practioners.

Antibiotic resistance is the ability of a microorganism to withstand the effects of antibiotics. It is

a specific type of drug resistance. Antibiotic resistance evolves via natural selection acting upon

random mutation, but it can also be engineered by applying an evolutionary stress on a

population. Once such a gene is generated, bacteria can then transfer the genetic information

in a horizontal fashion (between individuals) by plasmid exchange. If a bacterium carries several

resistance genes, it is called multiresistant or, informally, a superbug. The term antimicrobial

resistance is sometimes used to explicitly encompass organisms other than bacteria.

Antibiotic resistance can also be introduced artificially into a microorganism through

transformation protocols. This can aid in implanting artificial genes into the microorganism. If

the resistance gene is linked with the gene to be implanted, the antibiotic can be used to kill off

organisms that lack the new gene.

The researchers have investigated the anti-bacterial properties of various plants

attemting to develop a new antibiotic out of natural substances that could be used by

communities.

Birds nest fern is effective against infecting Escherichia Coli and Pseudomonas

aeruginosa. They content the potent antibiotic components like alkaloids, flavanoids, saponins

and tannins. They have strong defensive mechanism.

The study attempts to determine the potential of Bird’s nest fern extract by evaluating

the sensitivity of the Escherichia coli subjected to Kirby-Bauer Antibiotic Testing.

Objectives of the study

General objectives

The study generally aimed to determine the potential of bird’s nest fern extract as an

antibiotic for Escherichia Coli.

Specific objectives

Specifically the study aimed to:

1 Determine the sensitivity of E. coli to bird’s nest fern extract through Kirby-Bauer

antibiotic testing

2. Determine the treatment where the specimens of bacteria are most sensitive to

3. Determine the significant differences among treatments and replications

conducted.

Significance of the Study

Specialists estimate that at least 90% of antibiotic prescriptions are unnecessary or

inappropriate. Mutation of bacteria happens and occurs anytime by the transfer of resistance

genes from the other bacteria.

Perhaps a new revolution of antibiotic is declared to be a necessity.

The study “Antimicrobial Activity of Bird’s Nest Ferns Extract against Escherichia Coli

has a vital importance in determining the effective and cheap antibiotic out of commonly found

in the locality like bird’s nest fern which can also eradicate different kinds of bacteria, according

to past references. It is also significance in avoiding the use of harmful chemicals which can

pollute the air, kill other organism than the target pest and can destroy the biodiversity.

Hypotheses

H0: Bird’s Nest Ferns Extract has no antibacterial effect on Escherichia Coli,

H1: Bird’s Nest Ferns Extract has antibacterial effect on Escherichia Coli,

Scope and limitations of the Study

This study was limited only on the detection of the sensitivity of Escherichia

Coli to Bird’s Nest Fern extract and to the stated objectives of the research study.

This study was conducted at Sultan Kudarat Polytechnic State College- Laboratory High

School Research Center , EJC Montilla, Tacurong City from September 12-16 2009.

Definitions of Key Terms

Agar- is a gelatinous substance chiefly used as culture medium for microbiological work.

Antibiotic- used to kill bacteria that give different diseases.

Antimicrobial agent- the bacteria causing agent. Its the transfer of medium to medium.

Bird’s nest fern- is a tropical and sub-tropical plant which is native to the rain forests of Asia,

Africa, India , Polynesia and Australia .

Cefuroxime- is a second-generation cephalosporin antibiotic that has been widely available.

Escherichia coli- can cause serious food poisoning in humans, and are occasionally responsible

for costly product recalls.

Extract- comes from the leaves or any parts of a plant that were subjected to pounding and

extracting.

Incubation - The development of an infection from the time the pathogen enters the body until

signs or symptoms first appear.

Kirby-Bauer antibiotic testing (KB testing or disk diffusion antibiotic sensitivity testing)- is a

test which uses antibiotic-impregnated wafers to test whether particular bacteria are

susceptible to specific antibiotics.

Sterilization – a process used to ensure that the materials are safe and clean.

Zone of inhibition- This is an area around a paper disk or colony of bacteria or mold where no

other organisms are growing.

CHAPTER II

REVIEW OF RELATED LITERATURE AND STUDIES

Related Literature

Escherichia coli is a Gram negative bacterium that is commonly found in the lower

intestine of warm-blooded organisms (endotherms). Most E. coli strains are harmless, but

some, such as serotype O157:H7, can cause serious food poisoning in humans, and are

occasionally responsible for costly product recalls. The harmless strains are part of the normal

flora of the gut, and can benefit their hosts by producing vitamin K2, or by preventing the

establishment of pathogenic bacteria within the intestine.

E. coli are not always confined to the intestine, and their ability to survive for brief

periods outside the body makes them an ideal indicator organism to test environmental

samples for fecal contamination. The bacteria can also be grown easily and its genetics are

comparatively simple and easily-manipulated or duplicated through a process of metagenics,

making it one of the best-studied prokaryotic model organisms, and an important species in

biotechnology and microbiology.

A strain of E. coli is a sub-group within the species that has unique characteristics that

distinguish it from other E. coli strains. These differences are often detectable only on the

molecular level; however, they may result in changes to the physiology or lifecycle of the

bacterium. For example, a strain may gain pathogenic capacity, the ability to use a unique

carbon source, the ability to inhabit a particular ecological niche or the ability to resist

antimicrobial agents.

Different strains of E. coli are often host-specific, making it possible to determine the

source of fecal contamination in environmental samples.

For example, knowing which E. coli strains are present in a water sample allows to make

assumptions about whether the contamination originated from a human, another mammal or a

bird.

New strains of E. coli evolve through the natural biological process of mutation, and

some strains develop traits that can be harmful to a host animal. Although virulent strains

typically cause no more than a bout of diarrhea in healthy adult humans, particularly virulent

strains, such as O157:H7 or O111:B4, can cause serious illness or death in the elderly, the very

young or the immunocompromised.

Biology and biochemistry

Escherichia coli cells propel themselves with flagella (long, thin structures) arranged as

bundles that rotate counter-clockwise, generating torque to rotate the bacterium clockwise.E.

Coli is Gram-negative, facultative anaerobic and non-sporulating. Cells are typically rod-shaped

and are about 2 micrometres (μm) long and 0.5 μm in diameter, with a cell volume of 0.6 - 0.7

μm3. It can live on a wide variety of substrates. E. coli uses mixed-acid fermentation in

anaerobic conditions, producing lactate, succinate, ethanol, acetate and carbon dioxide. Since

many pathways in mixed-acid fermentation produce hydrogen gas, these pathways require the

levels of hydrogen to be low, as is the case when E. coli lives together with hydrogen-

consuming organisms such as methanogens or sulfate-reducing bacteria.

Optimal growth of E. coli occurs at 37°C but some laboratory strains can multiply at

temperatures of up to 49°C.

Growth can be driven by aerobic or anaerobic respiration, using a large variety of redox

pairs, including the oxidation of pyruvic acid, formic acid, hydrogen and amino acids, and the

reduction of substrates such as oxygen, nitrate, dimethyl sulfoxide and trimethylamine N-oxide.

The Bird's Nest Fern, Asplenium nidus, is a tropical and sub-tropical plant which is native

to the rain forests of Asia, Africa, India , Polynesia and Australia . In the wild, these largest of

epiphytes live up in the crowns of trees. Their means of nourishment is from nutrients found in

dust particles and rain which is collected in the plants deep, funnel-shaped rosette of leaves.

The genus name Asplenium (Spleenwort) is derived from the Greek words 'a' meaning 'not', and

'splen', meaning 'spleen' and refers to the medicinal properties the plant is supposed to have.

The specific epithet or species name 'nidus' means 'a nest', hence the common name Bird's

Nest Fern. The Bird's Nest Fern belongs to the 'Aspleniaceae' family under the fern order

'Polypodiales'.

The leathery, tongue-like, glossy, pale green, wavy-edged fronds of this fern are borne in

a tight rosette at the rhizome, forming a bowl-shaped clump with a fibrous center, which to

many resembles a bird's nest, or at least a seemingly sheltered spot for a bird's nest. Each frond

is entire or undivided and has a prominent, polished rib running down the center of the frond.

The new fronds uncurl from the hairy, dark scaled crown of the rosette in typical fern manner.

They are very soft and brittle when young and must not be handled.

As the new fronds grow in, the older ones gradually die. Mature fronds can grow to 3

feet in length and 8 inches in width, and under ideal conditions, a plant can reach 4 to 6 feet in

height, but, under normal home conditions the plant rarely exceeds 15 inches in height. The

stems of each frond are stumpy and erect. Another commonly grown variety is Asplenium

crispafolium, which has deeply ruffled fronds.

Phytochemical Content of Leaf Extract

Alkaloids are naturally occurring chemical compounds containing basic nitrogen atoms.

It protects plant from herbivores. They are anti-spasmodic, anti-

inflammatory,tranquilizer and vasodilator.

Flavonoids (or bioflavonoids), also collectively known as Vitamin P and citrin, are a class

of plant secondary metabolites. They are anti-allergic and antimicrobial and anti-

inflammatory.

Saponins, group of naturally occurring oily glycosides that foam freely when shaken with

water. They occur in a wide variety of plants, including acacia, soapwort, soaproot,

California pigweed, and many others. Saponins have been, and sometimes still are, used

as cleaning agents and as foam producers, notably in fire-extinguishing fluids. They have

a bitter taste

Tannins , group of naturally occurring oily glycosides that foam freely when shaken with

water. They are antimicrobial, protect against cancers and decrease blood lipids.

CHAPTER III

Research Methodology

This chapter discusses different mediums utilized in the research design and studies as

well the procedures and methods used in the conduct of the study. Also included are the

methods in gathering data as well as the statistical tools in the research.

Research Design

The study is experimental and comparative in nature. The study was conducted using

the Analysis of Variance (ANOVA) of Completely Randomized Design (CRD) with the three

treatments replicated three times in three trials with the same cultured bacteria per

replication. The treatments were as follows:

Treatment 1 -pure bird’s nest fern extract

Treatment 2 -5 ml bird’s nest fern extract + 5 ml water solution

Control - Cefuroxime

Material Gathering Procedure

The variables used in the study were bird’s nest fern and Escherichia coli. The

bacteria were collected from the urine oversaw by SKPSC Research Center for culturing

reasons. Bird’s nest fern were collected at SKPSC garden and were gathered fresh for this

contains more extract

Preparation of Experiment Set-Up

Escherichia Coli were cultured in the SKPSC Research Center for the study. The three

treatments, replicated thrice, were placed randomly on every Petri dish with the cultured

bacteria in it. Application of extract was administered by impregnating Bird’s Nest fern extract

unto sterile paper disks. Three trials were conducted for the validity for the study.

Materials and Equipment

The following were used for the conduct of the study:

For Extracting:

Digital weighing machine Mortar and pestle

Cheesecloth Graduated cylinder

Sterilized water Burner

Empty and clean bottle Bid’s Nest fern

Crucible tongs

For Sensitivity Testing:

Bunsen burner test tube

Inoculating loop incubator

Sterile cotton swab weighing scale

Petri dish Erlenmeyer flask

Forceps hot plate

Foil paper Auto clave

Sterile paper discs

Mueller-Huinton agar (2%)

Sodium chloride

Tetracycline impregnated filter-paper disc

Bacteria (Escherichia Coli)

McFarland Standard

Agar used for incubating organisms:

Blood agar

MacConky Agar

Methodology:

The following methods were used in the study:

Extraction of Bird’s Nest fern:

1. Collect the materials needed and prepare the setup.

2. Pound the 20 grams of bird’s nest leaves fern using the mortar and pestle

3. Squeeze the pounded leaves using the cheesecloth to obtain the extract.

4. Treatment 1 requires concentrated amount of bird’s nest fern extract and

requires no boiling. For the treatment 2, add 5 ml solution of pure bird’s nest fern

extract to 5 ml sterilized water. Dispense the mixture unto a test tube and set to

boiling water.

5. After obtaining the needed extract for both treatments, store solutions

securely unto sterilized empty bottles and seal.

Kirby-Bauer Antibiotic testing (direct method)

1. Growth for primary plate- inoculate into NSS

2. Streak to susceptible plate. E.coli MH with 2% NaCL

3. Let the inoculum dry for 3-5 minutes with the plates closed.

4. Placed the sterile paper disc impregnated with respective bird’s nest fern

extracts and cefuroxime(control) on the agar plate with sterile forceps. Press down

securely to ensure even contact.

-Disc must be equidistant with each other to allow zone of inhibition (ZI) in

millimeters

5. Incubate plates immediately or within 30 minutes for 24 hours at degrees

Celsius in an inverted position.

6. Read after 24-hour incubation.

Data Collection Procedure

Zone of Inhibition. After 24 hours, measure the diameter, in millimeters, of the region where

the Escherichia Coli were inhibited after 24 hours of exposure to the treatments. Zone of

inhibition will determine the remarks, whether the bacteria were resistant or sensitive to the

treatments. For the control, it was based on the antidote’s SIR legend (on hospital standards).

Positive (+) for ZI = sensitive

Negative (-) for ZI = resistant

Statistical Analysis of Data

The zone of inhibition of each replicates and the mean of the three replicates per

treatments were computed. The data gathered were used in answering the question: Which

treatment will give the best result in terms of antibacterial effect on Escherichia Coli after 24

hours?

Data gathered in this study was statistically analyzed using the Analysis of Variance of

Completely Randomized Design. The test for significant differences among treatments means is

subjected to Duncan‘s Multiple Range Test (DMRT)

CHAPTER IV

RESULTS AND DISCUSSION

This chapter presents the results of the tests conducted to determine the potential of

bird’s nest fern to be an antidote for Escherichia coli and the statistical analysis performed on

the data gathered. It is also presents the interpretations of the results of the analysis.

Zone of inhibitions of Escherichia coli 24 hours after contact to bird’s nest fern extract

for the different trials are being shown in the following table:

Table 1. Results of the sensitivity testing for the first trial.

Treatments R1 R2 R3 Total Mean Remarks

Zone of Inhibition

T1

(pure Bird’s Nest

fern extract)

55mm

54mm

56mm

165mm

55mm

SENSITIVE

T2

(5 ml Bird’s Nest

fern w/ 5 ml

H20)

0mm

2mm

0mm

2mm

0.67mm/

undefined

RESISTANT

Control

(Cefuroxime)

46mm

50mm

48mm

144mm

48mm

SENSITIVE

(SIR BASIS)


Negative (-) for ZI =resistant

Table1 shows that the result of the sensitivity test on Escherichia coli after 24 hours of

incubation and exposure to Bird’s Nest fern extract and the control group on its first trial.

Treatment 1 which is a pure Bird’s Nest fern extract has a mean zone of inhibition of 55

mm obtained from the zone of inhibition of its replicates: R1=55mm, R2=54mm and R3= 56mm.

treatment 2, a solution with bird’s nest fern plus water extract has a zone of inhibition of

0.67mm with replicates: R1=0mm, R2=2mm and R3=0mm. treatment 3 (Cefuroxime), the

control group, has a mean rate of 48 mm obtained from the mean rate of its replicates:

R1=46mm, R2=50mm and R3=48mm.

Based on the table 1, it was found out that Escherichia coli applied with treatment 1

got the highest zone of inhibition and remarked “sensitive”, which implies a good and effective

antibacterial effect. Moreover, it is not directly comparable to the control group since

Cefuroxime is manufactured for human dosage while Bird’s Nest fern extract intended for

direct inoculation to the bacteria. Conversely, treatment 2 (bird’s nest fern w/ H20) showed the

least ZI and therefore remarked as “resistant”.

Table 2. Results of the sensitivity for the second trial.

Treatments

R1 R2 R3

Total

Mean

RemarksZone Of Inhibition

T1

(pure bird’s nest

fern extract)

65mm

69mm

67mm

201mm

67mm

SENSITIVE

T2

(5 ml bird’s nest

fern w/ 5 ml H20)

0mm

0mm

0mm

0mm

0mm/

undefined

RESISTANT

Control

(Cefuroxime)

55mm

66mm

59mm

180mm

60mm

SENSITIVE

(SIR

BASIS)



Table2 shows that the result of the sensitivity test on Escherichia coli after 24 hours of

incubation and exposure to Bird’s Nest fern extract and the control group on its second trial.

Treatment 1 which is a pure bird’s nest fern extract has a mean zone of inhibition of 67mm

obtained from the zone of inhibition of its replicates: R1=65mm, R2=69mm and R3= 67mm.

Treatment 2, a solution with bird’s nest fern plus water extract has a zone of inhibition of 0mm

with replicates: R1=0mm, R2=0mm and R3=0mm. Treatment 3 (Cefuroxime), the control group,

has a mean rate of 60mm obtained from the mean rate of its replicates: R1=55mm, R2=66mm

and R3=59mm.




Cefuroxime is manufactured for human dosage while bird’s nest fern extract intended for direct

inoculation to the bacteria. Conversely, treatment 2 (bird’s nest fern w/ H20) showed the least

ZI and therefore remarked as “resistant.”

Table 3. Results of the sensitivity testing for the third trial.

Treatments

R1 R2 R3

Total

Mean


T1

(pure bird’s nest

fern extract)

55mm

56mm

58mm

169mm

56.34mm

SENSITIVE

T2

(5 ml bird’s nest

fern w/ 5 ml H20)

3mm

1mm

0mm

4mm

1.34mm

RESISTANT

Control

(Cefuroxime)

45mm 43mm 48mm 136mm 45.34mm SENSITIVE

(SIR BASIS)



Table 3 shows that the result of the sensitivity test on Escherichia coli after 24 hours of

incubation and exposure to Bird’s Nest fern extract and the control group on its third trial.

Treatment 1 which is a pure bird’s nest fern extract has a mean zone of inhibition of 56.34mm

obtained from the zone of inhibition of its replicates: R1=55mm, R2=56mm and R3= 58mm.

Treatment 2, a solution with bird’s nest fern plus water extract has a zone of inhibition of

1.34mm with replicates: R1=3mm, R2=1mm and R3=0mm. treatment 3 (Cefuroxime), the

control group, has a mean rate of 45.34mm obtained from the mean rate of its replicates:

R1=45mm, R2=43mm and R3=48mm.




Cefuroxime is manufactured for human dosage while bird’s nest fern extract intended for direct

inoculation to the bacteria. Conversely, treatment 2 (bird’s nest fern w/ H20) showed the least

ZI and therefore remarked as “resistant.”

Table 4. Analysis of Variance of the first trial.

Source of df SS MS F -value F -tab

variance 5% 1%

Replication 6.94 18.00

Treatment 6.94 18.00

Error

Total

**Significant

*Not Significant

Based on the analysis of variance, the computed Frequency value obtained for

replication is __.The tabular frequency (2,4)=6.94 at 0.5 and 18 at 0.1. Hence, the replication is

not significant. This means that replicates on the second trial do not really differ with each

other.

For treatment, the computed f-value obtained is ______ and the tabular F(2,4)=6.94 at

0.5level and 18 at 0.1 level of probability, the treatment is significant which means that

treatments really differ with each other in terms of zone of inhibition.

Table 5. Analysis of Variance of the second trial.

Source of

variance

df SS MS F -value F -tab

5% 1%



Error

Total

**Significant

*Not Significant



not significant. This means that replicates on the second trial do not really differ with each

other.




Table 6. Analysis of Variance of the third trial.

Source of

variance

df SS MS F -value F -tab

5% 1%



Error

Total

**Significant

*Not Significant



not significant. This means that replicates on the third trial do not really differ with each other.




Table 7. General tabulation of the trials conducted.

Treatments

R1 R2 R3

Total

Mean


T1

(pure bird’s nest

fern extract)

55mm

67mm

56.34mm

178.34mm

59.45mm

SENSITIVE

T2

(5 ml bird’s nest

fern w/ 5 ml

H20)

0.67mm

0mm

1.34mm

2.01mm

0.67mm

RESISTANT

Control

(Cefuroxime)

48mm

60mm

45.34mm

153.34mm

51.11mm

SENSITIVE

(SIR BASIS)



Table 7 shows the general tabulation of the trials conducted for the sensitivity of

Escherichia coli to different treatments used. Trials of treatment 1 which is a pure bird’s nest

fern extract has a mean zone of inhibition of 59.45mm obtained from the zone of inhibition of

its replicates: Trial1=55mm, Trial 2=67mm and Trial 3= 56.34mm. Treatment 2, a solution with

bird’s nest fern plus water extract has a zone of inhibition of 0.67mm with replicates: Trial

1=0.67, Trial 2=0mm and Trial 3=1.34mm.

Treatment 3 (Cefuroxime), the control group, has a mean zone of inhibition of 51.11mm

obtained from the mean of its trials: Trial 1=48mm, Trial 2=60mm and Trial 3=45.34mm.




Cefuroxime is manufactured for human dosage while Bird’s Nest fern extract is intended for

direct inoculation to the bacteria. Conversely, treatment 2 (bird’s nest fern w/ H20) showed

the least ZI and therefore remarked as “resistant.”

Table 8.General ANOVA

Source of

variance

df

SS

MS

F -value

F -tab

5% 1%



Error

Total

**Significant

*Not Significant

Based on the Analysis Of Variance, the computed Frequency value obtained for trial is

__.The tabular frequency (2,4)=6.94 at 0.5 and 18 at 0.1. Hence, the trial are not significant.

This means that result of the trial do not really differ with each other.



treatments really differ with each other in trials conducted in terms of zone of inhibition.

Statistically, the analysis of variance of the first, second and third trial showed almost

the same results. It therefore implies that the study is valid.

Results above showed high sensitivity of bacteria Escherichia coli to pure bird’s nest fern

extract as subjected to Kirby-Bauer Antibiotic Testing.

CHAPTER V

SUMMARY OF FINDINGS, CONCLUSION, AND RECOMMENDATION

This chapter discusses the findings, conclusions, and recommendations of the research

study on “Antimicrobial Activity of Bird’s Nest Ferns Extract Against Escherichia Coli.” It was

conducted at Sultan Kudarat Polytechnic State College- Laboratory High School Research Center

, EJC Montilla, Tacurong City from September 12-16 2009 using the Analysis of Variance of

Completely Randomized Design (CRD) with three treatments replicated three times.

*Major Finding

The major findings of this study were as follows:

1. Treatment 1(pure birds nest extract) got a general mean of its ZI with 59.45 mm while

Treatment 2 (5 ml bird’s nest extract + 5ml of sterilized water) got a general mean of

0.67mm.The control cefuroxime got a general mean of 51.11mm.

2. Escherichia coli showed high sensitivity when exposed to pure bird’s nest fern extract.

3. It is therefore found out that the treatment 1 is where Escherichia coli is most

sensitive to.

4. Based on the Analysis of Variance, treatments have significant differences among

each other in all trials conducted, while the replicates and trials are not significant

*Conclusions

The following conclusions were based from the major findings.

Based on the findings of the test conducted, bacteria Escherichia coli is favorably

sensitive to treatment 1, which is composed of pure birds nest fern extract from its leaves as it

remarked sensitive. The treatments showed highly significant differences while the replicates

and trials are very precise and showed not so much significance.

Likewise, the comparison between the control group and the treatments used cannot be

directly measured since the control (i.e.Cefuroxime) is manufactured considering the dosage for

human consumption while bird’s nest fern extract is made to combat E. coli in direct contact

with the bacteria itself.

Nevertheless, the study generally proved the great potential of bird’s nest fern extract

to be the radical antidote for the threatening public health nightmares, the E. coli.

*Recommendations

Studies on the different mediums had shown significant findings. In this connection,

the researchers strongly recommend that:

1. The sensitivity testing using pure birds nest extract to another sample of bacteria

be conducted.

2. Determine the SIR legend for birds nest fern extract to determine the accurate and

precise comparison with commercial antibiotics.

3. Further study should be conducted to manufactured birds nest fern in exact dosage

intended for human consumption.

Antimicrobial Activity of Bird’s Nest Fern Extract Against Escherichia Coli (an Experimental Research)

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