COMPARATIVE EFFECTIVENESS AND OPTIONAL PERIOD OF

THE FLOTATION METHOD USING NaCl, ZnSO4 AND MgSO4 FOR

THE DIAGNOSTIC OF SOIL-TRANSMITTED HELMINTHS

A SCIENTIFIC ARTICLE

RESEARCH REPORT

Submitted to fulfill the assignment and fit-out requisite to obtain

Bachelor of Medicine Degree

DWI WIDYANI ROSNIA SAVITRIE

22010110120017

UNDERGRADUATE PROGRAM BACHELOR OF MEDICINE

SCHOOL OF MEDICINE

DIPONEGORO UNIVERSITY

2014

ii

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RESEARCH AUTHENTICITY DECLARATION

The undersigned,

Name : Dwi Widyani Rosnia Savitrie

Student ID Number : 22010110120017

Address : Jl. Galang Sewu Raya no.1A, Semarang

Student Major in : Undergraduate Program in Medicine ( Bachelor of Medicine ) Faculty of

Medicine , Diponegoro University, Semarang.

Hereby declare that :

a) This scientific research article is based on my genuine idea, formula, and research, without

any help from other parties, except supervisor and known parties.

b) This scientific research article is authentic and has never been published for any academic

purpose in Diponegoro University or other university.

c) In this scientific research article there is no other people’s works or opinion which have been

used and published, unless there are clear acknowledgements of the original author names and

the original paper titles are included in the references.

Semarang, July 21st 2014

The declarant,

Dwi Widyani Rosnia Savitrie

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PREFACE

Alhamdulillah Thank God I praise to Almighty God, for the blessings and His grace that I

could finish these Academic Writing task. Writing the Academic Writing was done in order to fulfill

one of the requirements to achieve the Bachelor of Medicine Degree from Faculty of Medicine

Diponegoro University. I noticed it was very hard for me to finish this Academic Writing without help

and guidance from various quarters since the restructuring proposal until the complete report of the

results of this Scientific Writing. Together I deliver thanks and the highest appreciation to:

1. Prof. Soedharto P. Hadi, MES, PhD as the Rector of Diponegoro University Semarang

2. dr. Endang Ambarwati, SpKFR(K) as the Dean of Medicine

3. dr. Sudaryanto,M.Pd.Ked as the supervisor who had provided suggestions and thoughts

before and during this research

4. Mrs. Rahma, Mr. Sri Martono, Mr. Ngatimin, Mr. Inung for laboratory helps

5. My parents, Mum, Dad ,brother, and family who always gave material or moral supports

6. My friends who always supported in resolving this academic writing

7. And others we are not mentioned one by one on assistance directly or indirectly to this paper

can be solved with good

Finally, I hope that ALLAH SWT to recompense their kindness to everyone who had helped me.

Hopefully this academic writing will be useful for all of us.

Semarang, July 21st 2014

Dwi Widyani Rosnia Savitrie

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CONTENTS

Title ..................................................................................................................... i

Approval Page..................................................................................................... ii

Research Authenticity Declaration Page ............................................................. iii

Preface ....................................................................................................................iv

Contents ............................................................................................................... v

List of Tables ................................................................................................... viii

List of Figures .....................................................................................................ix

Abbreviations ...................................................................................................... x

Abstract...................................................................................................................xi

Abstrak...................................................................................................................xii

CHAPTER I INTRODUCTION ........................................................................... 1

1.1 Background of study ...................................................................................... 1

1.2 Research questions ......................................................................................... 5

1.3 Research purposes .......................................................................................... 5

1.3.1 General purposes ......................................................................................... 5

1.3.2 Special purposes .......................................................................................... 5

1.4 Research benefits ........................................................................................... 6

1.5 Research originality ........................................................................................ 7

CHAPTER II LITERATURE REVIEWS ............................................................. 8

2.1 Soil-transmitted helminths .............................................................................. 8

2.1.1 Ascaris lumbricoides (roundworm) .............................................................. 8

2.1.1.1 Morphology and life cycle of Ascaris lumbricoides .................................. 9

2.1.1.2 Pathology and clinical symptoms of Ascaris lumbricoides ...................... 11

2.1.2 Necator americanus and Ancylostoma duodenale (hookworm) .................. 11

2.1.2.1 Morphology and life cycle of hookworms ............................................... 12

2.1.2.2 Pathology and clinical symptoms of hookworms .................................... 14

2.1.3Trichuris trichiura (whipworm) .................................................................. 14

2.1.3.1 Morphology and life cycle of Trichuris trichiura ................................... 15

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2.1.3.2 Pathology and clinical symptoms of Trichuris trichiura .......................... 16

2.2 Diagnosis of soil-transmittedhelminths ......................................................... 17

2.2.1 Collected and preserved spesimen ............................................................ 17

2.2.2 Diagnostic techniques of soil transmitted helminth seggs ........................... 18

2.3 Flotation method .......................................................................................... 20

2.3.1 Flotation solution....................................................................................... 20

2.3.1.1 Sodium Chloride (NaCl) ......................................................................... 22

2.3.1.2 Magnesium Sulphate (MgSO4) ............................................................... 23

2.3.1.3 Zinc Sulphate (ZnSO4) ........................................................................... 25

2.3.2 Optional period.......................................................................................... 25

CHAPTER 3THEORITICAL FRAMEWORK, CONCEPTUAL

FRAMEWORK AND HYPOTHESES ............................................................... 27

3.1 Theoritical framework .................................................................................. 27

3.2 Conceptual framework ................................................................................. 28

3.3 Hypotheses ................................................................................................... 28

CHAPTER 4 RESEARCH METHODS ............................................................. 29

4.1 Research fields ............................................................................................. 29

4.2 Research locations and period ...................................................................... 29

4.3 Research design ............................................................................................ 29

4.4 Sample ......................................................................................................... 31

4.4.1 Inclusion criteria ........................................................................................ 31

4.4.2 Drop out criteria ........................................................................................ 32

4.4.3 Sampling method....................................................................................... 32

4.5 Research variables ........................................................................................ 32

4.5.1 Independent variable ................................................................................. 32

4.5.2 Dependent variable .................................................................................... 32

4.5.3 Confounding variable ................................................................................ 33

4.6 Operational definitions ................................................................................. 33

4.7 Data collection ............................................................................................. 34

4.7.1 Lab work materials .................................................................................... 34

4.7.2 Lab work equipment .................................................................................. 34

4.7.3 Type of data .............................................................................................. 35

4.7.4 Laboratory work ........................................................................................ 35

4.7.4.1 Collected and handling stool sample ....................................................... 35

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4.7.4.2 Making flotation solution ........................................................................ 37

4.7.4.2.1 Saturated Sodium Chloride solution (NaCl, s.g. 1.200) ........................ 37

4.7.4.2.2 Saturated Magnesium Sulfat Solution (MgSO4, s.g1200 ) .................... 37

4.7.4.2.3 Saturated Zinc Sulphate (ZnSO4-7H2O, s.g 1.200) .............................. 37

4.7.4.3 Flotation method..................................................................................... 38

4.8 Research protocol ......................................................................................... 40

4.9 Data analysis ................................................................................................ 41

4.10 Research ethics ........................................................................................... 42

4.11 Research schedule ...................................................................................... 42

CHAPTER V RESULTS.................................................................................... 43

5.1. Analysis of Samples .................................................................................... 43

5.1.1 Characteristic of research samples…………………………………...…….44

5.1.2 Prevalence of infection……………………………………………………..45

5.2. Analysis of Data .......................................................................................... 47

CHAPTER VI DISCUSSIONS .......................................................................... 56

CHAPTER VII CONCLUSIONS AND RECOMMENDATIONS ..................... 57

7.1. Conclusions ................................................................................................. 57

7.2. Recommendations ....................................................................................... 58

References ......................................................................................................... 59

Attachments ...................................................................................................... xiii

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LIST OF TABLES

Table 1. Research originality ................................................................................ 9

Table 2. Treatment of soil-transmitted helminths ................................................ 22

Table3. Classification intensity of worm infection .............................................. 27

Table4. Comparison of flotation solution............................................................ 29

Table5. The types of Magnesium Sulfate based on Hydrate contained ................ 31

Table 6.Research design ..................................................................................... 38

Table 7. Operational definitions ......................................................................... 41

Table 8. Research schedule ................................................................................ 49

Table 9. Characteristic of research subjects based on age……………………….52

Table 10. Characteristic of research subjects based on gender…………………..53

Table 11. Prevalence of Infection (in general)………………………………...…53

Table 12. Prevalence of infection based on gender………………………………54

Table 13. Prevalence of infection based on age………………………………….54

Table 14. Eggs counting number for each treatment groups…………………….56

Table 15. Normality test SaphiroWilk and Homegenity………………………...57

Table 16. Friedman test results…………………………………………………..61

Table 17. Wilcoxon test results…………………………………………………..62

Table 18. Difference test multivariate non paired Kruskal Wallis test…………..63

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LIST OF FIGURES

Figure 1.Ascarislumbricoidesadult male and female ........................................... 11

Figure 2.Fertile decorticated egg of Ascarislumbricoides.................................... 12

Figure 3. Infertile corticated egg of Ascarislumbricoides .................................... 12

Figure 4. Life cycle of Ascarislumbricoides ....................................................... 13

Figure 5.Necatoramericanusadult female ........................................................... 15

Figure 6. Hookworm egg .................................................................................... 16

Figure 7.Filariform larvae of hookworm ............................................................ 16

Figure 8. Life cycle of hookworms ..................................................................... 17

Figure 9.Trichuristrichiura adult male and female.............................................. 19

Figure 10.Trichuristrichiuraegg ......................................................................... 20

Figure 11. Adult maleTrichuristrichiura............................................................. 20

Figure 12.Life cycle of Trichuristrichiura .......................................................... 21

Figure 13.Theoritical framework ........................................................................ 35

Figure 14.Conceptual framework ....................................................................... 36

Figure 15.Research protocol framework ............................................................. 47

Figure 16. Box plot of flotation method for each solution in 15 minutes …….…58

Figure 17.Box plot of flotation method for each solution in 30 minutes …….…59

Figure 18.Box plot of flotation method for each solution in 45 minutes …….…59

Figure 19. Box plot of flotation method for each solution in 60 minutes …….…60

Figure 20. Box plot of flotation method for each solution in 70 minutes …….…60

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ABBREVIATIONS

EPG : Eggs per Gram

GIS : Geographical Information System

HCL : Hydrogen Chloride

MgSO4 : Magnesium Sulphate

NaCl : Natrium Chloride

PVA : Polyvinyl Alcohol

S.g : Specific Gravity

STH : Soil-Transmitted Helminths

WHO : World Health Organization

ZnSO4 : Zinc Sulphate

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ABSTRACT

Background. Factors that affect the effectiveness of flotation method in Soil-transmitted

helminths diagnostic are flotation solution and optional period. Further research is required to

determine the optional period that gives the best effectiveness from three flotation solutions,

which are NaCl, MgSO4, and ZnSO4.

Aim. To compare the effectiveness of NaCl, MgSO4 and ZnSO4 as flotation solution in

flotation method and the best optional period of each floating solution determined by

counting number of eggs.

Methods. This research was quasi-experimental analytic with two independent variable.

Independent variables 1 were flotation solutions (saturated NaCl, MgSO4 and ZnSO4) .

Independent variables 2 were optional period (15,30,45,60 and 70 minutes). Each

combination of treatments performed on 8 samples.

Results. There were significant differences between different of optional periods in one

solution .The same optional period for each solution ( saturated NaCl, saturated MgSO4 and

saturated ZnSO4) were not gives siginificant differences. The difference considered

significant when the p value < 0.05. The most number of eggs found in NaCl was at 60

minutes, ZnSO4 at 45 minutes, and MgSO4 at 45 minutes).

Conclusion. The optimum of optional period for each solution were: saturated NaCl (60

minutes), ZnSO4 (45 minutes), and MgSO4 (45 minutes). Flotation method using ZnSO4 as

solution with 45 minutes as optional period considered as the most effective method based on

number eggs found and time. In other hand, the effectiveness of the flotation method obtained

either with NaCl, MgSO4 and ZnSO4 in all level of optional period were same.

Keywords: flotation method, soil transmitted helminths, parasite diagnostic

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ABSTRAK

Latar belakang. Faktor-faktor yang mempengaruhi efektivitas pemeriksaan metode flotasi

dalam diagnosis cacing tanah yaitu jenis larutan flotasi dan periode opsional. Penelitian lebih

lanjut diperlukan untuk menentukan periode opsional yang memberikan hasil terbaik dari tiga

larutan flotasi yaitu larutan NaCl jenuh, MgSO4 jenuh, dan ZnSO4 jenuh.

Tujuan. Untuk membandingkan efektivitas NaCl, MgSO4 dan ZnSO4 sebagai larutan

pengapung dalam metode flotasi dan periode opsional terbaik untuk setiap larutan yang

digambarkan dengan menghitung jumlah telur.

Metode. Penelitian ini merupakan studi analitik quasi-eksperimental dengan dua variabel

bebas. Variabel bebas 1 adalah larutan flotasi (NaCl, MgSO4 dan ZnSO4). Variabel bebas 2

adalah periode opsional (15,30,45,60 dan 70 menit). Setiap kombinasi perlakuan dilakukan

pada 8 sampel.

Hasil. Terdapat perbedaan signifikan antara periode opsional berbeda dalam 1 larutan. Pada

periode opsional yang sama untuk semua larutan (NaCl,MgSO4 dan ZnSO4 ) tidak terdapat

perbedaan signifikan. Perbedaan bermakna signifikan bila nilai p < 0.05. Jumlah telur

terbanyak ditemukan pada NaCl menit ke 60, ZnSO4 menit ke 45, dan MgSO4 menit ke 45.

Simpulan. Periode opsional paling optimal untuk masing-masing larutan yaitu: NaCl (menit

ke 60), ZnSO4 (menit ke 45) and MgSO4 (menit ke 45). Metode flotasi menggunakan ZnSO4

jenuh sebagai larutan dengan 45 menit sebagai periode opsional memberikan hasil yang

paling efektif berdasarkan pada jumlah telur yang ditemukan dan waktu. Di sisi lain,

efektivitas metode flotasi yang diperoleh baik dengan larutan NaCl jenuh, MgSO4 jenuh dan

ZnSO4 jenuh dalam semua tingkat periode opsional adalah sama.

Kata kunci : metode flotasi, cacing tanah, diagnosis parasit