Antimicrobial Resistance for Enteric Pathogens Isolated from Acute

117 Pages·2011·1 MB·English

The Islamic University – Gaza
Deanship of Graduate Studies
Faculty of Science
Biological Sciences Master Program
Medical Technology

Antimicrobial Resistance for Enteric Pathogens Isolated from
Acute Gastroenteritis Patients in Gaza strip, Palestine.

By
Nahed Abdelateef

Supervisor

Dr. Abdelraouf A. Elmanama

Ph. D Microbiology

A Thesis Submitted in Partial Fulfillment of the Requirements for the
Degree of Master of Biological Sciences/ Medical Technology

May 2011
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Declaration

"I herby declare that this submission is my own work and that, to the best of
my knowledge and belief, it contains no material previously published or
written by another person nor material which to a substational extent has
been accepted for the award of any other degree of the university or other
institute, except where due acknowledgment has been made in the text".

Nahed Abdelateef

© Copyright by
Nahed Abdelateef
2011

Copyright.

All Right Reserved: No part of this work can be copied, translated or stored in
retrieval system, without prior permission of the author.

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DEDICATION

This thesis is dedicated to my family and
to my friends who supported me all the
way since the beginning of this study.

This thesis is also dedicated to all those
believe in the richness of learning.

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List of contents
Declaration ……………………………………………………….……….…….......... iv
Dedication ……………………………………………………………......................... v
List of figures………………………………………………………………..………... ix
List of tables ……………………………………………………………….................. x
List of abbreviations……………………………………………………….…..……… xi
Acknowledgments ……………………………………………………………............. xiii

CHAPTER I: INTRODUCTION
1.1 Overview ……………………….………..…………………………..................... 3
1.2 Objectives …………………………………………………………....................... 5
1.3 Significance ………………………………………………………………............ 5

CHAPTER II: LITERATURE REVIEW
2.1Acutegastroenteritis………………………………………………………..……….. 7
2.2 Antimicrobial resistance …………………………………………........................... 7
2.3 Diarrheal disease is the most common cause of illness ………………………...… 8
2.4 Common bacterial causes of diarrhea ……………...…………............................... 9
2.4.1 Salmonella spp history……………………….…………………….......…........... 9
2.4.2 Shigella history................................................................................…….……..... 10
2.4.3 Campylobacter history …………….…………...……………………..........….... 12
2.4.4 Yersinia enterocolitica history .………………….………………..............…..... 15
2.4.5 Aeromonas hydrophila history..………………………………….…................… 17
2.5 Epidemiology and antimicrobial resistance of gastroenteritis pathogen……..……. 19

CHAPTER III: MATERIALS AND METHODS
3.1 Materials ………………………………………………………………................. 28
3.1.1 Bacterial culture media…….………………………………………….…..... 28
3.1.2 Reagents ………………………………………………………...…….......... 28
3.1.3 Supplies ……………………………………………………..…………........ 29
3.1.4 Apparatus and equipments ………………………………….…………….... 29
3.2 Study area ……………………………………………………………………..….. 30
3.2.1 Samples …………………………………………………………………….. 30
3.2.2 Samples transport.………………………………………………...………… 30
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3.2.3 Questionnaires………………………………………………………………. 30
3.2.4 Permissions and ethical consideration ………………………...…………… 31
3.2.5 Data analysis …………………………………………………..…………… 31
3.3 Isolation and identification procedures of enteropathogenic bacteria………..…... 31
3.3.1 Salmonella and Shigella………………………………………..…...…….… 31
3.3.2 Campylobacter. …………………………………………………...…….….. 33
3.3.3 Yersinia enterocolitica ………………………………………...…………… 33
3.3.4 Aeromonas hydrophila ………………………………………...………..….. 33
3.4 Antimicrobial susceptibility for the bacterial isolates…………….…...………….. 34
3.5 Staining and biochemical tests……………………………………...………… ..... 34
3.5.1 Gram stain …………………………………………………………...…....... 34
3.5.2 Oxidase test …………………...……………………………………............. 35
3.5.3 Catalase test…………………………………………………..….................. 35
3.5.4 API 20 E test (BioMerieux, France)…………………………...…................ 35
3.5.5 API campy test……………………………………………………...………. 35
3.6 Specific antisera………………………………………………............................... 36
3.6.1 Serotyping of Shigella spp……………………………….............................. 36
3.6.2 Serotyping of Salmonella spp……………………......................................... 36

CHAPTER IV: RESULTS
4.1 Description of study samples…………...………………………………………… 37
4.1.1 Distribution of the samples in terms of residence area of the study samples.. 37
4.1.2 Sex and age distribution of the study samples ……………...………..…...... 38
4.1.3 Socioeconomic status of the study samples ……………………...……….... 38
4.2 Isolation and identification of enteropathogenic bacteria……………...…..…....... 39
4.2.1 Salmonella and Shigella…………………………………............................... 39
4.2.2 Campylobacter ................................................................................................ 40
4.2.3 Yersinia enterocolitica ………………………………………...……………. 41
4.2.4 Aeromonas hydrophila ………………………………………...………......... 42
4.2.5 Number and types of isolated enteropathogenic bacteria among population.. 43
4.2.6 Isolated enteropathogenic bacteria distributed by residence…….…………... 43
4.2.7 Occurrence of enteropathogenic bacteria among the different age groups….. 44
4.2.8 Distribution of enteropathogenic bacteria according to gender……..…...….. 45
4.2.9 Clinical symptoms in relation to presence of isolated bacteria…………....… 46
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4.2.10 Correlation of the socioeconomic status of the study samples with isolated
enteropathogens………………………………………………..…...……… 47
4.3 Antimicrobial susceptibility profile…………………………..………………....... 48
4.3.1 Antimicrobial profile of enteropathogenic bacteria isolated from patient...... 48
4.3.2 Prescription of antibiotics to the diarrheal patients…………….…................ 50
4.4 Correlation of socioeconomic factors with diarrhea………………….…...……... 51
4.4.1 Correlation between diarrhea and domestic animals in the houses….....…... 51
4.4.2 Correlation between diarrhea and some furniture in the house…….........…. 52
4.4.3 Correlation between diarrhea and current water access in the houses…….... 52
4.4.4 Correlation between diarrhea and drinking water in the house…....……...... 53
4.4.5 Correlation between diarrhea and sewage disposal…………...…………..... 54
4.5 Clinical signs and symptoms of diarrheal patients……………….…...…...……... 55
4.6 Management and follow-up of diarrheal patients ……………….….......……...… 56

CHAPTER V: DISCUSSION
5.1 Children age less than 5 years old are more susceptible to infection…………….. 58
5.2 Diarrhea and enteropathogenic bacteria is more frequent in crowded houses with
low room's number………………………………………………………………... 5 9
5.3 Diarrhea is more frequent in houses rearing poultry, sheeps, and pigeons……..... 59
5.4 Improvement in sewage disposal is vital in the reduction of diarrheal diseases..... 60
5.5 Correlation between water access, drinking water and diarrhea……….…..…...... 61
5.6 Sign and symptoms of diarrheal patients…………..………………….…….......... 63
5.7 Follow-up and convalescence of diarrheal patients………………………...…...... 64
5.8 Prevalence of bacterial enteropathogens……………………………….…...…...... 65
5.9 Distribution of isolated enteropathogenic bacteria by residence……….…...……. 65
5.10 Antimicrobial resistance for enteropathogenic bacteria……………....………… 67
5.11 Dehydration caused by diarrhea………………….…………………..…………. 68
5.12 Relation of the weather with isolation rate of enteropathogenic bacteria….......... 70

CHAPTER VI: CONCLUSIONS AND RECOMMENDATIONS
6.1 Conclusions ……………………………………………………………………….. 71
6.2 Recommendations………………………………………….…………………...…. 73
REFERENCES……………………………………………………….………....…..... 75
ANNEXES…………………………………………………………………..….……... 100
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List of figures

Figure (1.2): Distribution of causes of death among children……...…………. 8

Figure (1.3): Isolation and identification procedures of enteropathogenic

bacteria………………………………………………………….. 3 2

Figure (4.1): Colony morphology for Salmonella…………………………….. 39

Figure (4.2): API 20E results for isolated bacteria……………………..……... 40

Figure (4.3): Agglutination with specific antisera for Salmonella ……............ 40
.
Figure (4.4): Colony morphology for Campylobacter spp…………...…....….. 40

Figure (4.5): (Modified CCDA) incubated in microerophilic condition

at 42°C…………………………………………………………... 41

Figure (4.6): Identification of Campylobacter spp. by API campy test………. 41

Figure (4.7): Colony morphology for Yersinia enterocolitica on Yersinia

selective agar……………………………………………………. 41

Figure (4.8): Identification of Yersinia enterocolitica by API 20E………..… 42

Figure (4.9): Colony morphology for Aeromonas hydrophila…………...….... 42

Figure (4.10): Identification of A. hydrophila by API 20E…………................ 42

Figure (4.11): Enteropathogenic bacteria isolated from the samples……......... 43

Figure (5.1): Levels of dehydration in children with acute diarrhea…….......... 69

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List of table

Table (4.1): Primary health care centres included in this study………............. 34

Table (4.2): Distribution of the cases and controls due to sex and age……..... 38

Table (4.3): Distribution of the cases and controls due to Socioeconomic

status……………………………………………………………... 39

Table (4.4): The isolated enteropathogenic bacteria by residence……………. 44

Table (4.5): Enteropathogenic bacteria among the different age group............ 45

Table (4.6): Enteropathogenic bacteria among the gender…………................ 45

Table (4.7): Clinical symptoms in relation to presence of enteropathogens..... 46

Table (4.8): Socioeconomic status of the study samples with isolated

enteropathogenic bacteria…........................................................... 47

Table (4.9): Antimicrobial susceptibility of isolated enteropathogenic

bacteria…………………………………………………………... 49

Table (4.10): Prescription of antibiotics to the diarrheal patients…….............. 50

Table (4.11): Domestic animals in the houses and diarrhea….......................... 51

Table (4.12): Correlation between diarrhea and some furniture in the house... 52

Table (4.13): Current water access in the houses….......................................... 53

Table (4.14): Drinking water in the houses……............................................... 54

Table (4.15): Sewage disposal and diarrhea……………………................... 55

Table (4.16): Clinical signs and symptoms of diarrheal patients…….............. 56

Table (4.17): Management and follow-up of diarrheal patients……............. 57

Table (5.1): Clinical features of infection with selected diarrheal pathogens... 64

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