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Chapter 1
Introduction to the Study
Chapter one, consists of five parts, namely: (1)
Background and Theoretical Framework of the Study, (2)
Statement of the Problem and Hypothesis, (3) Significance
of the Study, (4) Definition of Terms, and (5) Delimitation
of the Study.
Part One, Background and Theoretical Framework of the
study, gives the reasons for the choice of the problem and
the variables considered in the conduct of the study.
Part Two, statement of the Problem and Hypothesis,
describes the problem and its purpose in conducting the
study and the hypothesis to be tested.
Part Three, Significance of the Study, enumerates the
benefits that may be derived from the results of the study
and persons who would benefit from them.
Part Four, Definition of Terms, deals with the
conceptual and the operational definitions of important
terms used in the study.
Part Five, Delimitation of the Study, sets the limit
and scope of the study.
of N 010 35.00
a typical
to about
young coral
30-40
island
meters elevation
100 meters.
rising
proceeding
the enteric bacteria, which are facultatively anaerobic gramnegative rods that live in the intestinal tracts of animals in
health and disease (Todar, 2008).
The science of antibiotics has remained and will
remain for many years, one of the most interesting natural
sciences, in both theoretical and practical aspects. Microbial
natural products still appear as the most promising source of
the future antibiotics that society is expecting (Pelaez,
2006).
For the past years, pathogens underwent mutation which
enabled them to resist antimicrobials, thereby threatening
millions of people worldwide. Thus, it is imperative to screen
more and more bacteria from different soil samples for
antimicrobial activity in hope of getting some bacterial strains
that produce antibiotics that have not been discovered yet and
active against drug resistant pathogens.
Dependent Variables:
Bacterial Isolates in
Brgy. Sinapsapan,
Jordan, Guimaras in
the high area
Bacterial Isolates in
Brgy. Sinapsapan,
Jordan, Guimaras in
the intermediate area
Bacterial Isolates in
Brgy. Sinapsapan,
Jordan, Guimaras in
the low area
Zone of inhibition
on Staphylococcus
aureus BIOTECH
1582
Zone of Inhibition
on Escherichia
coli BIOTECH 1634
Negative Control:
distilled water
Definition of Terms
The following terms are defined to ensure clarity and
understanding of the study:
Antibacterial effect- According to Mozo (2008), it is an
effect that is destructive to bacteria or suppressing their
reproduction or growth, effective against bacterial
infections.
In this study, antibacterial effect refers to the action
of screened bacterial isolates against S. aureus and E. coli.
10
11
12
Chapter 2
Review of Related Literature
This chapter presents the conceptual literature regarding
this study to support the background for greater understanding
of the research outcome. This is consisted of the following:
(1)Staphylococcus aureus morphology, (2) its pathogenicity,
(3)Escherichia colis morphology, (4) colony characteristics,
(5) its pathogenicity, (6) soil, and Barangay Sinapsapan, Jordan
Guimaras.
Staphylococcus aureus
Morphology. According to Brooks, Carroll, Butel, Morse and
Mietzner (2010) staphylocci are gram positive spherical cells,
usually arranged in grapelike irregular clusters. They grow
readily on many types of media and are active metabolically,
fermenting carbohydrates and producing pigments that vary from
white to deep yellow.
13
14
15
16
17
18
19
20
21
22
23
24
Soil
According to Alanis (2005) serious infections caused by
bacteria that have become resistant to commonly used antibiotics
have become a major global healthcare problem in the 21st
century. Staphylococcus aureus, for instance, a virulent
pathogen that is responsible for a wide range of infections
including pimples, pneumonia, osteomyelitis, endocarditis and
bacteremia, has developed resistance to most classes of
antibiotics. For more than two decades, clinicians and public
health officials have faced hospital acquired methicillinresistant Staphylococcus aureus (MRSA), which also bears
resistance too many antibiotics. During much of this time,
vancomycin has been the therapeutic answer to MRSA, but that
paradigm has changed. Vancomycin resistant strains have emerged
clinically. Vancomycin-resistant S. aureus (VRSA) challenges
clinicians, not only because of vancomycin and methicillin
resistance, but also because of resistance to many other
antibiotics, including aminoglycosides, macrolides, and
fluoroquinolones. Fortunately, newer therapeutic agents,
daptomycin, linezolid, and a streptogramin combination
(quinupristin/dalfopristin) have entered the clinical arena in
the past few years. However, certain undesirable side effects
25
26
27
28
of N 010 35.00
E 122 30.47.
rising rapidly
proceeding inland
a typical
to about
young coral
30-40
islands
meters elevation
100 meters.
with the
when
Going
29
ridges.
According to Espaola (Personal Communication, October 10,
2013) many areas of the barangay are still untouched and
unexplored due to the accessibility. The barangay is composed of
different soil colors. It varies from dark brown and reddishyellowish brown. The texture of the soil is mostly coarse. With
regards to its structure, the arrangement of the soil particles is
in different aggregates. Soil granules clump or bind together.
According to Eckerwall (Personal Communication, October 10,
2013) in the valleys
found
but
the ridges
coral rocks
and
corn is planted.
larger trees.
Rocks
reach
some
The
10 cm
offers
little
can be
with sharp
mainly
fruit and
of
a possibility to
and
30
Chapter 3
Research Design and Methodology
This chapter consists of Three (3) parts: Part One
Purpose of the Study and Research Design, Part Two Method, and
Part Three Data Analysis Procedure.
Part One, Purpose of the Study and Research Design,
elucidates on the purpose of the study and describes the
research design.
Part Two, Method, the instrument used, and the procedures
employed in gathering the data pertinent to the study.
Part Three, Statistical Data Analyses, explains the
statistical tools employed in the treatment of data and the
quantitative part which includes the measurement of the zone of
inhibition.
31
32
Chemicals used
The following chemicals were prepared and used for this
study: Nutrient broth, Nutrient Agar, McFarland 0.5, Normal Saline
Solution, mineral oil, safranin, grams iodine, crystal violet,
Fluconazole and 70% Alcohol (as disinfectant).
Procedures
Sterilization of Materials. The glassware were washed,
dried and wrapped with a newspaper. They were placed inside the
autoclave at 15 psi for 15-20 minutes at 121C. After
sterilization, they were dried in a drying oven. The materials
were stored for later use.
Sampling Site Description. The sampling sites are located
in Barangay Sinapsapan,
on the lands
E 122 30.47.
N 010
121
33
0C
34
were taken and spread evenly over the surface of nutrient agar
35
36
37
0C.
( )
Where
X = diameter of the petri dish used (90 mm)
Y = diameter of the punched hole on the agar plate (10 mm)
Z = number of descriptive scale used (4)
38
Description
0-20 mm
inactive;
21-40 mm
partially active
41-60 mm
Active
61-80 mm
very active
39
40
Chapter 4
Results and Discussion
41
Table 2
Bacterial isolates obtained in the three sampling sites of Brgy.
Sinapsapan, Jordan, Guimaras.
Sampling Site
Number of Isolates
High Area
Intermediate Area
Low Area
42
Table 3
Characteristics of Bacterial Isolates in the High, Intermediate
and Low Area Sampling Sites of Brgy. Sinapsapan, Jordan,Guimaras
Isolate Number
Sampling site
H1
High
H2
High
H3
High
H4
High
H5
High
H6
High
I1
Intermediate
I2
Intermediate
I3
Intermediate
I4
Intermediate
I5
Intermediate
I6
Intermediate
L1
Low
L2
Low
L3
Low
L4
Low
L5
Low
L6
Low
Colony Characteristics
Irregular; Flat; Undulate; Dull;
Opaque; Non- pigmented; Smooth
Circular; Flat; Entire; Dull; Opaque;
Non- pigmented; Smooth
Punctiform; Flat; Entire; Dull;
Transluscent;
Nonpigmented;
Smooth
Rhizoid; Flat; Filamentous; Dull;
Transluscent;
Nonpigmented;
Smooth
Circular; Flat; Entire; Dull; Opaque;
Yellow; Smooth
Circular; Flat; Entire; Dull; Opaque,
Orange; Smooth
Rhizoid; Flat;Filamentous; Dull;
Transluscent;
Nonpigmented;
Smooth
Circular; Flat; Entire; Dull; Opaque;
Non- pigmented; Smooth
Punctiform; Flat; Entire; Dull;
Transluscent;
Nonpigmented;
Smooth
Irregular; Flat; Undulate; Dull;
Opaque; Non- pigmented; Smooth
Filamentous; Flat; Filamentous;
Dull; Opaque; Non- pigmented;
Smooth
Circular; Flat; Entire; Dull; Opaque;
Yellow; Smooth
Rhizoid;
Flat;Filamentous;
Dull;Opaque;
Non- pigmented;
Smooth
Filamentous; Flat; Filamentous;
Dull; Opaque; Non- pigmented;
Smooth
Punctiform; Flat; Entire; Dull;
Transluscent;
Nonpigmented;
Smooth
Irregular; Flat; Undulate; Dull;
Transluscent;
Nonpigmented;
Smooth
Circular; Flat; Entire; Dull; Opaque;
Yellow; Smooth
Circular; Flat; Entire; Dull; Opaque;
Orange; Smooth
Gram
Reaction
Bacilli in clusters
Cocci in clusters
Cocci in chain
Cocci in clusters
Diplobacilli
Staining
Cocci in clusters
Cocci in clusters
Cocci in clusters
Cocci in clusters
Cocci in clusters
43
60
44
Active
50
isolate 1
40
Partially Active
isolate 2
isolate 3
30
isolate 4
isolate 5
Inactive
20
isolate 6
Antibiotic
10
Negative Control
0
24 hours
48 hours
72 hours
45
Active
50
isolate 1
Partially Active
40
isolate 2
isolate 3
30
isolate 4
Inactive
20
isolate 5
isolate 6
10
Antibiotic
0
24 hours
48 hours
72 hours
Negative
Control
46
60
50
isolate 1
Patially
Active
40
isolate 2
isolate 3
30
isolate 4
Inactive
20
isolate 5
isolate 6
10
Antibiotic
Negative
Control
24 hours
48 hours
72 hours
47
40
35
isolate 1
30
isolate 2
25
isolate 3
Inactive
20
isolate 4
15
isolate 5
10
isolate 6
Antibiotic
Negative
Control
24 hours
48 hours
72 hours
1634
48
40
Partially Active
35
30
isolate 1
isolate 2
25
isolate 3
Inactive
20
isolate 4
isolate 5
15
isolate 6
10
Antibiotic
Negative Control
5
0
24 hours
48 hours
72 hours
49
40
Partially Active
35
isolate 1
30
isolate 2
25
isolate 3
20
Inactive
isolate 4
15
isolate 5
10
isolate 6
Antibiotic
0
24 hours
48 hours
72 hours
Negative Control
50
51
Table 4
Analysis of Variance with Repeated Measures for the Zone of
Inhibition of Bacterial Isolates in the Different Sampling Sites
against Staphylococcus aureus BIOTECH 1582.
Effect
Time*Isolate
Wilks
lambda
Value
Hypothesi
s df
Error
df
Signifi
cance
Partial
Eta
Squared
0.825
1.868
74
0.125
0.092
52
Table 5
Analysis of Variance with Repeated Measures for the Zone of
Inhibition of Bacterial Isolates in the Different Sampling Sites
against Escherichia coli BIOTECH 1634.
Effect
Wilks
lambda
Value
Hypothes
is df
Error
df
Signifi
cance
Partial
Eta
Squared
Time*Isola
te
Wilks
lambda
0.681
3.915
74
0.006*
0.175
53
Discussion
A total of 18 bacterial isolates in the 3 sampling sites
with variable colony and cell characteristics in each sampling
site were collected. The bacterial isolates are all flat, dull,
non-pigmented and smooth. I1 is a diplobacilli while the rest
are
either
clustered
or
occur
singly.
Amongst
the
different
bacteria
are
divided
into
three
groups
viz
Cocci
and
influence
Spirilla
the
in
soil.
microbial
The
major
population,
soil
factors
distribution
and
which
their
and
type
of
organism
but
also
their
activities.
also
to
microorganisms
in
soil
and
thereby
54
proliferate
the
Seasonal
changes
in
soil
temperature
affect
microbial
chief
source
of
energy
and
food
for
most
of
the
soil
matter
influence
directly
or
indirectly
on
the
Ecological
relationships
among
soil
organisms
Gram
positive,
aerobic
or
facultative
endospore
55
fixer
(Liu
et
al.,
2006),
denitrifiers
(Suharti
et
al.,
general,
the
soil
bacterial
isolates
exhibited
an
isolates
may
be
producers
of
novel
bioactive
have
been
aminoglycosides,
macrolides,
isolated
anthracyclines,
nucleosides,
polyketides,
and
characterized
glycopeptides,
peptides,
actionomycins,
and
betalactams,
polyenes,
tetracyclines
including
polyester,
(PrashithKekuda,
are
extracellular
metabolites
which
are
normally
agents,
(Kekuda,
gram-positive
drugs,
Shobha,
rods
&
grow
immunoregulators
Onkarappa,
as
2010).
filaments,
and
antiparasitic
Actinomycetes
branching
rods,
are
and
56
I1
Actinomycetes
has
as
almost
parallel
gram-positive,
characteristics
filamentous
and
to
diplobacilli.
substances
2005). They
that
can
may
have
produce
IAA,
antibacterial
property
siderophore,
phytase,
and
solubilized
various
sources
of
organic
and
to
Sharma
et
al.
(2001),
the
ability
of
is
influenced
by
environmental
conditions
such
as
etc.
one
species
will
be
more
efficient
than
the
others and will fill the niche with its offspring leaving no
57
58
Chapter 5
Summary, Conclusions, Implications, and Recommendations
Chapter Five consists of four parts: (1) Summary of the
Findings, (2) Conclusions, (3) Implications, and (4)
Recommendations.
Part One, Summary of the findings; state the salient points
of the study.
Part Two, Conclusions, presents the interferences drawn
from the results of the study.
Part Three, Implications, clarifies the relationships
between the present investigation and the theories presented in
relation to the antibacterial screening of soil bacterial
isolates from Sinapsapan, Jordan, Guimaras against Escherichia
coli and Staphylococcus aureus.
Part Four, Recommendations, offer some suggestions in terms
of the findings, conclusions, and recommendations.
59
60
Conclusions
Based on the experimental study, the results showed the
following:
1. There were 6 soil bacterial isolates obtained in the
high, intermediate, and low sampling sites in Brgy. Sinapsapan,
Jordan, Guimaras.
2. Based on the characterization of the bacterial isolates
in Brgy. Sinapsapan, Jordan, Guimaras, the shared
61
62
Implications
This study was conducted to screen the antibacterial
properties of soil bacterial isolates from Sinapsapan, Jordan,
Guimaras against S. aureus and E. coli. The findings have shown
that different isolates are more effective on S. aureus than on
E. coli. It is therefore possible for the isolates to be
considered as a prospective source of antibiotic against on S.
aureus than E. coli. Searching for new possible source of
antibiotics is necessary because it becomes a manner for
bacteria to be resistant to certain antibiotics. Antibiotic
resistant bacteria are increasing nowadays in number that makes
the treatment for some diseases to be difficult.
Bacteria become resistant more quickly when antibiotics are
used improperly. Since there are kinds of bacteria that are
63
64
Recommendations
Based on the conclusions of the study, the following are
recommended:
1. The study was limited only on the characterization of
the colonies and gram staining technique, it is further
recommended to do special staining methods such as flagellar
staining to determine the arrangement of the flagella, endospore
staining to determine whether the isolates are spore-former and
65
66
67
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Appendix A
Sampling Site Map
74
75
Appendix B
Characteristics of Bacterial Isolates in the High Area Sampling
Site of Brgy. Sinapsapan, Jordan Guimaras.
76
Appendix C
Characteristics of Bacterial Isolates in the Intermediate Area
Sampling Site of Brgy. Sinapsapan, Jordan Guimaras.
77
Appendix D
Characteristics of Bacterial Isolates in the Low Area Sampling
Site of Brgy. Sinapsapan, Jordan Guimaras.
Appendix E
Antibacterial Data Sheet
Sampling Site: ____HIGH______ Test Pathogen:____E. coli______
78
79
80
81
82
83
84
Appendix F
Least Significant Difference for Staphylococcus aureus Multiple
Comparisons
95% Confidence Interval
(I) Group
Isolate 1
(J) Group
Isolate 2
Isolate 3
Mean Difference
(I-J)
-.2067
Std. Error
.91919
Sig.
.823
Lower
Bound
-2.0675
Upper
Bound
1.6541
-1.2574
.91919
.179
-3.1182
.6034
Isolate 4
-1.3422
.91919
.152
-3.2030
.5186
Isolate 5
-.5404
.91919
.560
-2.4012
1.3204
Isolate 6
1.6200
.91919
.086
-.2408
3.4808
-18.4530(*)
1.29993
.000
-21.0845
-15.8214
.2067
.91919
.823
-1.6541
2.0675
Isolate 4
Isolate 5
-1.0507
-1.1356
-.3337
.91919
.91919
.91919
.260
.224
.719
-2.9115
-2.9964
-2.1945
.8101
.7252
1.5271
Isolate 6
1.8267
.91919
.054
-.0341
3.6875
-18.2463(*)
1.29993
.000
-20.8779
-15.6147
Isolate 1
1.2574
.91919
.179
-.6034
Isolate 2
1.0507
.91919
.260
-.8101
2.9115
Isolate 4
-.0848
.91919
.927
-1.9456
1.7760
Isolate 5
.7170
2.8774(*)
.91919
.91919
.440
.003
-1.1438
1.0166
2.5778
4.7382
-17.1956(*)
1.29993
.000
-19.8271
-14.5640
Isolate 1
1.3422
.91919
.152
-.5186
Isolate 2
1.1356
.91919
.224
-.7252
2.9964
Isolate 3
.0848
.91919
.927
-1.7760
1.9456
Isolate 5
.8019
.91919
.388
-1.0589
2.6627
Isolate 6
2.9622(*)
.91919
.003
1.1014
4.8230
-17.1107(*)
1.29993
.000
-19.7423
-14.4792
Isolate 2
Isolate 3
Positive
Control
Isolate 3
Isolate 6
Positive
Control
Isolate 4
Positive
Control
Isolate 5
Positive
Control
Isolate 1
Isolate 1
.5404
.91919
.560
-1.3204
Isolate 2
.3337
.91919
.719
-1.5271
2.1945
Isolate 3
-.7170
.91919
.440
-2.5778
1.1438
Isolate 4
-.8019
.91919
.388
-2.6627
1.0589
Isolate 6
2.1604(*)
.91919
.024
.2996
4.0212
-17.9126(*)
1.29993
.000
-20.5442
-15.2810
Isolate 1
-1.6200
.91919
.086
-3.4808
-1.8267
.91919
.054
-3.6875
.0341
Positive
Control
Isolate 6
Isolate 2
Isolate 3
-2.8774(*)
.91919
.003
-4.7382
-1.0166
Isolate 4
-2.9622(*)
.91919
.003
-4.8230
-1.1014
Isolate 5
-2.1604(*)
.91919
.024
-4.0212
-.2996
-20.0730(*)
1.29993
.000
-22.7045
-17.4414
Positive
Control
Positive
Control
Isolate 2
Isolate 1
18.4530(*)
1.29993
.000
15.8214
18.2463(*)
1.29993
.000
15.6147
20.8779
Isolate 3
17.1956(*)
1.29993
.000
14.5640
19.8271
Isolate 4
17.1107(*)
1.29993
.000
14.4792
19.7423
17.9126(*)
1.29993
.000
15.2810
20.5442
20.0730(*)
1.29993
.000
17.4414
22.7045
Isolate 5
Isolate 6
3.1182
3.2030
2.4012
.2408
21.0845
85
Appendix G
Least Significant Difference for Escherichia coli Multiple
Comparisons
95% Confidence Interval
(I) Group
Isolate 1
(J) Group
Isolate 2
isolate 3
Mean
Difference
(I-J)
.0863
Std.
Error
.80336
Sig.
.915
Lower Bound
-1.5400
Upper Bound
1.7126
-.7004
.80336
.389
-2.3267
.9260
isolate 4
-.7833
.80336
.336
-2.4097
.8430
isolate 5
.8626
.80336
.290
-.7637
2.4889
isolate 6
.9619
.80336
.239
-.6645
2.5882
Antibiotic
-4.3559(*)
1.13613
.000
-6.6559
-2.0560
Isolate 2
Isolate 1
-.0863
.80336
.915
-1.7126
1.5400
isolate 3
-.7867
-.8696
.80336
.80336
.334
.286
-2.4130
-2.4960
.8397
.7567
isolate 4
isolate 5
.7763
.80336
.340
-.8500
2.4026
isolate 6
.8756
.80336
.283
-.7508
2.5019
Antibiotic
1.13613
.000
-6.7422
-2.1423
isolate 3
-4.4422(*)
Isolate 1
.7004
.80336
.389
-.9260
Isolate 2
.7867
.80336
.334
-.8397
2.4130
isolate 4
-.0830
.80336
.918
-1.7093
1.5434
isolate 5
.80336
.80336
1.13613
.059
.045
.003
-.0634
.0359
-5.9555
3.1893
3.2885
-1.3556
isolate 4
1.5630
1.6622(*)
-3.6556(*)
Isolate 1
.7833
.80336
.336
-.8430
Isolate 2
.8696
.80336
.286
-.7567
2.4960
isolate 3
.0830
.80336
.918
-1.5434
1.7093
isolate 5
1.6459(*)
.80336
.047
.0196
3.2723
isolate 6
1.7452(*)
.80336
.036
.1189
3.3715
Antibiotic
-3.5726(*)
1.13613
.003
-5.8726
-1.2726
isolate 5
Isolate 2
Isolate 1
-.7763
-.8626
.80336
.80336
.340
.290
-2.4026
-2.4889
.8500
isolate 3
-1.5630
.80336
.059
-3.1893
.0634
isolate 4
-1.6459(*)
.80336
.047
-3.2723
-.0196
isolate 6
.0993
.80336
.902
-1.5271
1.7256
Antibiotic
-5.2185(*)
Isolate 1
1.13613
.000
-7.5185
-2.9185
-.9619
.80336
.239
-2.5882
isolate 3
isolate 4
-.8756
-1.6622(*)
-1.7452(*)
.80336
.80336
.80336
.283
.045
.036
-2.5019
-3.2885
-3.3715
.7508
-.0359
-.1189
isolate 5
-.0993
.80336
.902
-1.7256
1.5271
Antibiotic
-5.3178(*)
1.13613
.000
-7.6177
-3.0178
Antibiotic
Isolate 1
4.3559(*)
1.13613
.000
2.0560
Isolate 2
4.4422(*)
1.13613
.000
2.1423
6.7422
isolate 3
3.6556(*)
1.13613
.003
1.3556
5.9555
isolate 4
3.5726(*)
5.2185(*)
5.3178(*)
1.13613
1.13613
1.13613
.003
.000
.000
1.2726
2.9185
3.0178
5.8726
7.5185
7.6177
isolate 6
Antibiotic
isolate 6
Isolate 2
isolate 5
isolate 6
2.3267
2.4097
.7637
.6645
6.6559
86
Appendix H
Zone of inhibition data of S. aureus
Number of
trials and its
24 HOURS
48 HOURS
72 HOURS
corresponding
(mm)
(mm)
(mm)
replicates
1-1
52
55
59
1-2
31
38
38
1-3
20
61
74
2-1
37
50
55
2-2
22
48
51
2-3
20
48
53
3-1
25
62
65
3-2
25
35
43
3-3
25
50
54
87
Appendix I
Zone of Inhibition data of E.coli
Number of
trials and its
24 HOURS
48 HOURS
72 HOURS
corresponding
(mm)
(mm)
(mm)
replicates
1-1
25
32
38
1-2
20
26
33
1-3
27
35
46
2-1
20
22
35
2-2
19
24
36
2-3
26
31
41
3-1
21
24
29
3-2
18
42
42
3-3
24
37
30
Appendix J
Descriptive Statistics for Staphylococcus aureus
Zone of Inhibition: High Land
88
89
90
Appendix K
Descriptive Statistics for Escherichia coli
Zone of Inhibition: High Land
91
92
93
Appendix L
Picture of Zone of inhibition
94
95
Appendix M
Letter to UPLB Biotech
April 8, 2014
DR. REYNALDO V. EBORA
BIOTECH Director
PNCM-BIOTECH, UPLB
Los Banos, Laguna 4031
Dr. Ebora:
Good day Sir!
I would like to request for the purchase of activated cultures
of Staphylococcus aureus and Escherichia coli that will be used
by my undergraduate advisees, Ma. Sophia Estella C. Tajolosa and
Kristine Joy R. Estrella for their antibacterial research study.
Aside from my supervision, our Biology department has also a
registered microbiologist to guide and assist them in handling
these bacteria. Our microbiology lab is also provided with the
necessary equipment and gadgets in order to qualify and fit for
a Biosafety level 2 organisms manipulation.
Attached herewith are the accomplished forms needed for our
request and for your perusal. I look forward for your favorable
acceptance of my request for the success of my students
undergraduate research study as part of their chosen career
path. Thank you and more power.
Very yours truly,
REY G. TANTIADO
Biological Science Instructor and Research Adviser
Bio. Sci., CAS, West Visayas State University
96
Appendix N
Letter of Permission
April 29, 2014
MR. AUDIE A. SULADAY
Head
Central Laboratory Section
West Visayas State University
Dear Mr. Suladay:
Greetings!
We, Ma. Sophia Estella C. Tajolosa and Kristine Joy R. Estrella,
incoming fourth year students of BS Biology will be conducting a research
experiment as a partial fulfillment of the requirement in our course subject,
Thesis Writing in Biological Sciences (BIO 230A).
In line with this, we would like to request the following chemicals and
laboratory apparatus for our thesis entitled Antibacterial Screening of Soil
Bacterial Isolates from Sinapsapan, Jordan Guimaras against Escherichia coli
and Staphyloccocus aureus this May 2014 at RH 104.
Attached hereof is the list of the following apparatus and chemicals
that we are to use in the said study.
We are looking forward for the granting of our request.
Thank you very much.
Respectfully Yours,
______________________________
MA. SOPHIA ESTELLA C. TAJOLOSA
Researcher
_____________________________
KRISTINE JOY R. ESTRELLA
Researcher
Noted by:
______________________
PROF. REY G. TANTIADO
Thesis Adviser
_____________________
DR. NANCY S. SURMIEDA
Dean, College of Arts and Sciences
_______________________
MR. AUDIE A. SULADAY
In Charge, Central Science Laboratory
97
98
99
100
101
102
Appendix X: Incubation
103
104
105
106
RESEARCH CERTIFICATION
This is to certify that KRISTINE JOY R. ESTRELLA and MA.
SOPHIA ESTELLA C. TAJOLOSA have submitted to me their manuscript
entitiled ANTIBACTERIAL SCREENING OF SOIL BACTERIAL ISOLATES
FROM SINAPSAPAN, JORDAN, GUIMARAS AGAINST Escherichia coli
AND Staphylococcus aureus.
I have examined the same and found it in order.
PRENCY C. YERRO M. A.
Student Research Coordinator
________________
Date
________________
Date
________________
Date
________________
Date
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Date