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)
Positive (+) for ZI = sensitive
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)
Positive (+) for ZI = sensitive
Negative (-) for ZI =resistant
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.
Based on the table 2, 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 3. Results of the sensitivity testing for the third trial.
Treatments
R1 R2 R3
Total
Mean
RemarksZone Of Inhibition
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)
Positive (+) for ZI = sensitive
Negative (-) for ZI =resistant
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.
Based on the table 3, 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 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%
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 6. Analysis of Variance of the third trial.
Source of
variance
df SS MS F -value F -tab
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 third 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 7. General tabulation of the trials conducted.
Treatments
R1 R2 R3
Total
Mean
RemarksZone Of Inhibition
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)
Positive (+) for ZI = sensitive
Negative (-) for ZI =resistant
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.
Based on the table 7, 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 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%
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 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.
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 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.