CAGAYAN STATE UNIVERSITY COLLEGE OF ARTS AND SCIENCES

CHARACTERIZATION OF AIRBORNE DRUG RESISTANT BACTERIA IN

SOLID WASTE DUMPING SITE IN TUGUEGARAO CITY, CAGAYAN

A Thesis
Presented to the
Faculty of College of Arts and Sciences
Cagayan State University
Carig Sur, Tuguegaro City

In Partial Fulfilment
of the Requirements for the Degree of
Bachelor of Science in Biology

JANNIE CEL PASCUAL AGGASID

LORLYN FLORES CATULIN

June 2017

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 CAGAYAN STATE UNIVERSITY COLLEGE OF ARTS AND SCIENCES

APPROVAL SHEET

In partial fulfillment of the requirements the degree, Bachelor of Science in

Biology this thesis entitled, “Characterization of Airborne Drug Resistance

Bacteria from Solid Dumping Site of Tuguegarao City, Cagayan”, prepared and

submitted by Jannie Cel P. Aggasid and Lorlyn F. Catulin, has been examined and

recommended for oral defense.

May 12, 2017 LIRIO GUDINA C. MANGAWIL, Ph.D.

Date Adviser

Approved by the committee on oral defense with the grade of ________%.

ANDY L. CATULIN, Ph.D.

Chairman

JEROME TAMAYAO, MST MADEL B. TANGUILAN, MAB

Member Member

PATRIANNE PADUA
English Critique

Accepted and approved in partial fulfillment of the requirements for the Degree in
Bachelor of Science in Biology

__________________ JANE SAMBRANA, Ph.D.

Date College Dean

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DEDICATION

To My
PAPA AND MAMA
Sisters
Who stood by me through all the years......
Who patiently sailed with me in my journey
To my DREAMS......
This is for you. -E

To The
Angel of My Life
DADDY
My Beloved Mother
To My Brothers and Sister
For their unfading Love and Support
You are always there guiding my road to
SUCCESS......
This is for you. -L

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ACKNOWLEDGEMENT

The researchers wishes to give due recognition, appreciation and gratitude to the

numerous persons who generously gave their time, effort, encouragement and help in

the completion of this study.

Our warm and very inadequate thanks to Dr. Lirio Gudina C. Mangawil, our

thesis adviser, for being patient, unselfish and for sharing her expertise in giving

suggestions and recommendations for the accomplishment of this study;

To the Program Coordinator and ever supportive Chairman of the Panel, Sir Andy

Catulin, for his undeniable passion in helping the researchers organize this book

properly;

To the panelists, Madam Unnis and Sir Jerome, for their comments and

suggestions for the improvement of the paper;

To Dr. Theresa B. Dimalanta, our college dean, for being supportive and letting

us conduct our study;

To Sir Frederick Realiza, our statistician who patiently computed our data for

free;

To Ma’am Patrianne Padua, our English critique, for patiently edited our paper for

free;

To Sir Crizaldy Binarao, our statistician who enlightened us with explanations;

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To the Panelists of Campus Wide Search for Undergraduate Thesis, Doc Gerly,

Doc Virgie and Madam Sylvia, for commendations and bountiful knowledge that

you’ve shared during the defense.

To Ma’am Gladys and Ma’am Hannah, the Laboratory Technician of CSU, for

allowing the researchers to use the laboratory equipments and facilities;

To Dr. Julius Capili, dean of CAHS, for comments and suggestions to improve

the paper;

To CVMC- Ethics Board Committee, for sharing their knowledge and expertise in

proper protocol;

To our supportive instructors: Madam Madel Batang and Madam Pauline

Ibanez, for being there for us since thesis proposal;

To all the staff of DOST - RSTL, Madam Junica, Madam Lara and Sir Arjay,

for letting us use their laboratory media and equipment for free;

To CENRO – Tuguegrarao headed by Atty. Mora and the personnel-in-charge of

the dumpsite for their assistance during the collection of sample;

To the staff of DA – RADDL, Madam Maychelle for the assistance; Dr.Jeff

Bassig for helping us to complete proper equipment used in Media Prep;

To the staff of DOH – MDR TB Lab, Madam Rhea for assisting us throughout

the experiment;

To the Office of the Mayor – Hon. Bienvenido de Guzman, for his warm

financial assistance given to materialize this study;

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To the compassionate special people in our hearts, Christian and Mark, for being

there for us whenever we needed assistance and for the sacrifices you have done just

to support us on this study;

Without the help of these personalities, the completion and success of this study

would have not been possible.

God, Almighty Father, whose compassion, eternal love unending protection,

guidance, wisdom and the highness and greatness of all.

Thank you.

Ecelyn

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TABLE OF CONTENTS

Page

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

APPROVAL SHEET……………………………………………...............ii

DEDICATION........................................................................….................iii

ACKNOWLEDGEMENT.…………………………..…………...............iv

TABLE OF CONTENTS……….…………………….….……..…..........vii

LIST OF TABLES.......................................................................................x

LIST OF FIGURES ....................................................................................xi

ABSTRACT……....…………………………….…..………......…...........xii

CHAPTER

I. INTRODUCTION

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

Objectives of the Study …….................….....……..........................3

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

Scope and Delimitation ……………………....................................4

Time and Locale of the Study ………...............................................5

II. REVIEW OF RELATED LITERATURE

Multiple-drug Resistant Bacteria .....................................................6

Types of MDR Infections.................................................................10

Suspected Drug Resistant Microorganisms .....................................12

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Staphylococcus aureus ............................................................12

Enterococcus spp. ....................................................................13

Streptococccus pneumoniae .....................................................14

Enterobacteriaceae Family ......................................................15

Pathogenicity ......................................................................................16

Antibiotics ..........................................................................................18

Test Antibiotics ...............................................................................19

Penicillin ...................................................................................19

Ampicillin .................................................................................20

Oxacillin ....................................................................................21

Erythromycin ............................................................................21

Tetracycline ..............................................................................22

Rifampin ...................................................................................23

Vancomycin...............................................................................24

Linezolid ...................................................................................25

Related Studies .................................................................................28

III. METHODOLOGY

Materials ............................................................................................33

Sampling Site......................................................................................33

Preparation of Agar ............................................................................34

Collection of Sample...........................................................................36

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Replica Plate Method.........................................................................36

Kirby-Bauer Susceptibility Test .......................................................37

Confirmatory test: Inoculation of Presumed Unidentified Bacteria..38

Confirmatory Test: Gram Staining Technique .................................39

Statistical Analysis ...........................................................................39

IV. RESULTS AND DISCUSSION

Total Population Count of Colonies per Station in the Dumpsite ....40

Microbial Isolates from Selective Medium........................................41

Cultural and Morphological Characteristics of Bacterial Isolates.....44

Zone of Inhibition and Antibiotic Susceptibility of Microbial

Isolates...............................................................................................47

Antibiotic Resistance Pattern of Bacterial Isolates............................49

V. SUMMARY, CONCLUSIONS AND RECOMMENDATIONS

Summary ..........................................................................................51

Conclusion .......................................................................................53

Recommendation..............................................................................53

APPENDICES

A. Acceptance Letter.........................................................................54

B. CENRO Permit.............................................................................55

C. DOH Communication Letter.........................................................56

D. Ethics Clearance ...........................................................................57

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E. Map of Dumpsite..........................................................................58

F. Isolates to Selective Media............................................................59

G. Microscopic View of the Isolates..................................................60

H. Kirby-Bauer Disc Diffusion ..........................................................61

I. Raw Data ........................................................................................62

J. Documentation ...............................................................................64

REFERENCES.............................................................................................70

CURICULUM VITAE.................................................................................66

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List of Tables

Table

1.1a Mean of Microbial Population (CFU)............................................33

1.1b Single Factor ANOVA of Total Microbial Population....................33

1.2a Cultural Characteristics of Colonies Isolated.................................36

1.2b Morphological Characteristics of Colonies Isolated.......................37

1.3 Mean of Zone of Inhibition (mm) and Susceptibility Bacterial Isolates

found in the dumpsite.....................................................................................39

1.4 Antibiotic Resistance Pattern of bacterial isolates from the

dumpsite.........................................................................................................40

A Confirmation of Bacterial Isolates on Selective Media...................54

1.5 Raw Data of Zone of Inhibition of Isolates.....................................57

1.6 Classification through Protocol Testing...........................................58

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List of Figures

Figure

A Dumpsite Map (Aerial View).............................................48

B Dumpsite Map (Station View)............................................48

C Station 1..............................................................................50

D Station 2..............................................................................50

E Station 3..............................................................................50

F Panoramic View of Dumpsite.............................................51

G Medical Wastes on Dumpsite..............................................51

H Unsegregated Wastes on Dumpsite......................................51

I Microscopic View of Suspected Isolates.............................55

J Zone of Inhibition................................................................56

1 Open Plate Method...............................................................59

2 Researcher on Collecting Site..............................................59

3 Selective Media for Replica Plating.....................................59

4 Replica Plating Method.........................................................59

5-6 Researchers conducting Streak Plate Method......................60

7-10 Researchers conducting Gram-Staining Technique……..................60

11-15 Antibiotic Susceptibility Testing......................................................61

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ABSTRACT

TITLE : CHARACTERIZATION OF AIRBORNE

DRUG RESISTANT BACTERIA IN

SOLID WASTE DUMPING SITE IN

TUGUEGARAO CITY, CAGAYAN

RESEARCHERS : JANNIE CEL P. AGGASID

LORLYN F. CATULIN

DEGREE : BACHELOR OF SCIENCE IN BIOLOGY

YEAR : 2017

INSTITUTION : CAGAYAN STATE UNIVERSITY

CARIG CAMPUS, TUGUEGARAO CITY

ADVISER : LIRIO GUDINA C. MANGAWIL, Ph.D.

This study entitled “CHARACTERIZATION OF AIRBORNE DRUG

RESISTANT BACTERIA IN SOLID WASTE DUMPING SITE IN

TUGUEGARAO CITY, CAGAYAN” was conducted to detect the occurrence of

antibiotic resistant bacteria.

The emergence of drug resistant bacteria in the community is one of the

major threat in public health worldwide. The study was conducted to determine the

air microbial population and detect the presence of airborne drug-resistant bacteria in

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the Solid Waste Dumping site of Tuguegarao City, Cagayan. Highest microbial

population was observed in sampling station 2 which was located nearer to the

ground. Using replica plating, six (6) isolates which were screened using selective

media were confirmed to be E. coli (MACS1E-A), S. aureus ..(BPAS2F-B),

Streptococcus sp. (BAPS2A-C), Klebsiella sp. (MACS3E-D), Salmonella sp.

(MACS2E-E), and Shigella sp. (MACS2F-F) through standard microbiological

procedures.

The antibiotic susceptibility profile of the isolates was determined using

conventional assay. All isolates were tested in eight (8) different classes of

antibiotics. Results on zone of inhibition were compared with natural antibiotic

profile, which is the standard of the National Committee for Clinical Laboratory

Standards (NCCLS). Isolate MACS2F-F revealed the highest resistance as it shows

100% resistance to eight selected antibiotics. Both isolates of MACS1E-A and

MACS2E-E showed resistance to 7 out of the 8 antibiotics (87.5%). Isolate

MACS3E-D showed resistance to 4 out of the 8 antibiotics (50%). Gram-positive

isolates BPAS2A-B and BAPS2A-C showed low resistance to the eight selected

antibiotics. BPAS2A-B was resistant to 3 out of the 8 antibiotics (37.5%) and

BAPS2A-C showed 0% resistant to the eight selected antibiotics.

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