CO-INFECTION OF SCHISTOSOMA HAEMATOBIUM AND ESCHERICHIA COLI IN PUPILS ATTENDING SOME SELECTED SCHOOLS, NIGERIA

ABSTRACT

This study was designed to investigate co-infection of Schistosoma haematobium and Escherichia coli in pupils attending some selected primary schools in Zaria, Nigeria. A total of 520 urine samples were collected from the pupils and were analyzed for ova of Schistosoma haematobium and also the urines were also cultured for Escherichia coli. The ova of Schsitosomiasis was determined using the standard centrifugation method. E,coli was isolated using Cystiene Lactose Electrolyte Deficient (CLED) Agar and was subcultured on Eosin Methylene Blue(EMB) Agar. IMVIC biochemical test were carried out on the isolates. Further biochemical test was carried out to confirm the isolates were Escherichia coli. Antibiotic susceptibility test was carried out on the E,coli isolate using disk diffusion method. The prevalence of Schistosoma haematobium was 20.4%, Escherichia coli was 4.6% and the co-infection was 4.6%. The prevalence rates of Schistosoma haematobium infection in LEA Z, LEA J, LEA G, and LEA P were 14.6%, 26.9%, 18.5% and 21.5% respectively. The prevalence of E. coli and co-infection in LEA Z, LEA J, LEA G and LEA P were 2.3%. 5.4%, 4.6% and 6.2%. The highest prevalence of Schistosoma haematobium was found among the age group of 10-14years There was a significant association (χ²=8.936, df=3, P=0.030*) between age and Schistosoma haematobium infection. The prevalence of E.coli was found to be higher among the age group of 15-19 years and the prevalence was higher in the female (4.7%) than male (4.6%). There was no significant association between E. coli infection and age. The prevalence of Schistosoma haematobium in male pupils (23.5%) was found to be higher than the female pupils (15.0%). There was significant association between Schistosoma haematobium and Gender. The prevalence of Escherichia coli in females pupils was higher when compared to the male pupils. There was no significant association between E. coli and gender. Higher prevalence of

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Schistosoma haematobium was found among pupils who use streams /lakes as source of drinking water. There was significant association between S. haematobium and source of drinking water.The antibacterial susceptibility profile of the isolates was determined using 10 different antibiotics. All the isolates were susceptible to Gentamicin. Some of the isolates were resistant to

Ciprofloxacillin, Amoxycillin, Chloramphenicol, Tetracycline, Sulphamethoxazole Trimethropin, Nalixidic acid, Ampicillin, Doxycycline and Nitrofurantoin.

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TABLE OF CONTENTS

Cover Page…………………………………………………………………………………………i

Fly Leaf……………………………………………………………………………………………ii

Title Page…………………………………………………………………………………………iii

Declaration………………………………………………………………………………………..iv

Certification……………………………………………………………………………………….v

Dedication………………………………………………………………………………………...vi

Acknowledgements………………………………………………………………………………vii

Abstract………………………………………………………………………………………….viii

Table of Contents………………………………………………………………………………….x

List of Tables…………………………………………………………………………………….xv

List of Figures…………………………………………………………………………………...xvi

List of Appendices……………………………………………………………………………...xvii

CHAPTER ONE

1.0         INTRODUCTION……………………………………………………………………….1

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

1.2         Statement of the Research Problem…………………………………………………….3

1.3         Justification of the Research…………………………………………………………….4

1.4         Aim………………………………………………………………………………………..5

1.5         Objectives………………………………………………………………………………....5

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CHAPTER TWO

2.0         LITERATURE REVIEW……………………………………………………………….6

2.1         Urinary Tract Infections………………………………………………………………...6

2.2         Clinical Manifestations of Urinary Tract Infection……………………………………6

2.2.1     Asymptomatic bacteriuria…………………………………………………………………7

2.2.2     Cystitis…………………………………………………………………………………….7

2.2.3     Acute pyelonephritis………………………………………………………………………8

2.3         Escherichia coli…………………………………………………………………………...8

2.4         Classification/Taxonomy………………………………………………………………...9

2.5         Habitat…………………………………………………………………………………..10

2.6         Cell Structure and Physiology…………………………………………………………11

2.7         Pathogenesis of Escherichia coli……………………………………………………….11

2.8         Enteric/diarrhoea E. coli……………………………………………………………….11

2.8.1     Enteropathogenic E. coli…………………………………………………………………12

2.8.2     EnterohaemorrhagicE. coli……………………………………………………………....12

2.8.3     Enterotoxigenic E. coli…………………………………………………………………...13

2.8.4     Enteroaggregative E. coli………………………………………………………………...13

2.8.5     Enteroinvasive E. coli……………………………………………………………………14

2.8.6    Diffused adherent E. coli………………………………………………………………...14

2.9         Uropathogenic E. coli…………………………………………………………………...14

2.9.1    Virulence factors…………………………………………………………………………15

2.9.2     Mannose-sensitive adhesins……………………………………………………………...19

2.9.3     Toxin genes………………………………………………………………………………23

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2.10      Taxonomy of Shistosoma haematobium ………………………………………………26

2.11      Structure………………………………………………………………………………...27

2.12      Life cycle and biology of the worm…………………………………………………….27

2.13      Transmission and Risk Factors for Infection…………………………………………30

2.14      Persistence, Latency, and Natural History of Infection……………………………...30

2.14.1 Persistence………………………………………………………………………………..30

2.14.2 Latency and Natural History……………………………………………………………..31

CHAPTER THREE

3.0         MATERIALS AND METHODS………………………………………………………32

3.1         Study Area………………………………………………………………………………32

3.2         Sample Size……………………………………………………………………………...32

3.3         Ethical Approval………………………………………………………………………..33

3.4         Administration of Questionnaire………………………………………………………33

3.5         Sample Collection……………………………………………………………………….33

3.6         Laboratoty Examination of Urine Sample………………………………………………..33

3.6.1     Examination for the Ova of Schistosoma haematobium………………………………………….33

3.6.2      Preparation of Media for the Growth of Escherichia coli…………………………………………………..34

3.7         Cultivation of Escherichia coli…………………………………....................................35

3.7.1     Isolation of E. coli………………………………………………………………………..35

3.7.2     Gram staining…………………………………………………………………………….36

3.7.3     Biochemical Characterization of E. coli…………………………………………………37

3.8         Confirmation of E. coli using Microgen™ GnA-ID Kit……………………………...37

3.9         Antibiotic Susceptibility Testing……………………………………………………….38

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3.9.1     Measurement of diameter of the inhibition zone………………………………………...39

3.9.2    Multi-drug resistance index……………………………………………………………...39

3.10      Statistical Analysis……………………………………………………………………...39

CHAPTER FOUR

4.0         RESULTS……………………………………………………………………………….40

4.1         Age Factor and Gender Related Prevalence of Schistosoma haematobium …….......40

4.2        Test of Association of Risk Factors in relation to Schistosoma haematobium………40

4.3        Test of Association of Symptoms in relation to Schistosoma haematobium…………44

4.4        Biochemical Characterization of Escherichia coli…………………………………….46

4.5        Age Factor and Gender Related Prevalence of Escherichia coli……………………..46

4.6     Test of Association of Risk Factors In relation to Escherichia coli …………………..46

4.7      Test of Association of Symptoms In relation to Escherichia coli ……………………..50

4.8         Age Factor and Gender Related to Co-infection of Schistosoma haematobium

and Escherichia coli ……………………………………………………………………52

4.9         Test of Association of Risk Factors In relation to Co-infection with

Schistosoma haematobium and Escherichia coli.……………………………………...52

4.10      Test of Association of Symptoms In relation to Co-infection with

Schistosoma haematobium and Escherichia coli ……………………………………...56

4.11     Prevalence of Schistosoma haematobium, Escherichia coli and Co-infection

of Schistosoma haematobium and Escherichia coli. …………………………………..60

4.12     Prevalence of Schistosoma haematobium, Escherichia coli and Co-infection

of Schistosoma haematobium and Escherichia coli In relation to Locations.……….60

4.13     Antibiotic Susceptibility of Escherichia coli Isolated From Urine…………………..63

4.14      Multiple Antibiotic Resistance Phenotype of Escherichia coli Isolated from Urine..63

4.15     Multiple Antibiotic Resistance Indices of Escherichia coli Isolated from Urine…...63

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CHAPTER FIVE

5.0         DISCUSSION…………………………………………………………………………...67

CHAPTER SIX

6.0         CONCLUSION AND RECOMMENDATION…………………………………….....72

6.2         Conclusion………………………………………………………………………………72

6.3         Recommendations………………………………………………………………………72

REFERENCES………………………………………………………………………………….74

APPENDICES…………………………………………………………………………………..94

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LIST OF APPENDICES

Appendix                                                                          Title                                                                                      Page

I                     Questionnaire…………………………………………………………………………….94

II                  Ethical Consent…………………………………………………………………………..95

III                 Letter of Introduction………………………………………………………………….....96

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CHAPTER ONE

1.0                                                                          INTRODUCTION

1.1         Background of the Study

Urinary tract infections (UTI) represent one of the most common infections encountered in medical practice today and occurring from the neonate to the geriatric age group (Kunin, 1994). Despite the widespread availability of antibiotics, UTI remains the most common bacterial infection in the human population (Tambekar et al., 2006). About one hundred and fifty million individuals had been reported to be affected by UTIs annually worldwide (Gupta et al., 2001). Urinary tract infections occur as a result of the microbial colonization of urine and the invasion of any structure of the urinary tract by microbial organisms such as bacteria, viruses, and/or parasites (Stamm, 1999; Stamm, 2008).

Urinary tract infections associated with Schistosoma haematobium affects the entire genitourinary tract (Ifeanyi et al., 2009). Bacterial infections are often recurrent and important in the prepatent period of urinary schistosomiasis which may be instrumental in precipitating renal failure (Ifeanyi et al, 2009). In urinary schistosomiasis, secondary bacterial infections are common and in men can involve the seminal vesicles, spermatic cord, and to a lesser extent, the prostate. In women, infection can involve the cervix and fallopian tubes and can cause infertility (Ifeanyi et al., 2009).

Urinary tract infection (UTI) is the commonest microbial infectious disease in community practice with a high rate of morbidity and financial cost. Urinary tract infections are described as bacteriuria with urinary symptoms. Urinary tract infection can affect lower and sometimes both lower and upper urinary tracts. The term cystitis had been used to define the lower UTI infection

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and is characterized by symptoms such as dysuria, frequency, urgency, and suprapubic tenderness. The presence of the lower UTI symptoms does not exclude the upper UTI which is often present in most UTI cases (Sobel and Kaye, 2010). The treatment of UTI can be classified into uncomplicated and complicated on the basis of their choice of treatment (Sabra and Abdel-Fattah, 2012). Urinary Tract Infection is more common in females than in males as female urethra structurally found less effective for preventing the bacterial entry (Warren et al., 1999). It may be due to the proximity of the genital tract and urethra and adherence of urothelial mucosa to the mucopolysaccharide lining (Schaeffer et al., 2001: Akortha and Ibadin, 2008). The other main factors which make females more prone to UTI are pregnancy and sexual activity (Arul et al., 2012). In pregnancy, the physiological increase in plasma volume and decrease in urine concentration develop glycosuria in up to 70% women which ultimately leads to bacterial growth in urine (Lucas and Cunningham 1993). Sexual activity in females also increases the risk of urethra contamination as the bacteria could be pushed into the urethra during sexual intercourse as well as bacteria being massaged up the urethra into the bladder during child birth (Ebie et al., 2001: Kolawole et al.,2009).

The resulting disease conditions from UTI include cystitis and pyelonephritis which is known to be non-age discriminatory as it affects both older persons and infants. Moreover, pyuria as evidenced by the inflammation of the genitourinary tract is common in subjects with asymptomatic bacteriuria (Nicolle et al., 2005). Asymptomatic UTI in particular has been associated with an increased risk of developing pyelonephritis, maternal and infant morbidity, pre-term labour and low birth weight (Oyagade et al., 2004). Urinary tract infections of both bacterial and parasitic origins had been associated with high incidence of squamous cell carcinoma of the bladder and the cervix (Schwartz, 1984); Escherichia coli and Schistosoma spp.

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are the most widely reported UTI-caused by bacteria and parasite respectively (Ariyo et al.,2004).

Microbial invasion being the basis of urinary tract infection could be seen in various clinical manifestations resulting in various disease conditions in both male and female of all ages. Young adults particularly female are, however, the most at risk of bacteriuria. For example, in the United States, UTIs result in approximately 8 million physician visits and more than 100,000 hospital admissions per year of sexually active women treated annually for UTIs. Up to 95% of the UTI cases in the U.S are treated with antibiotics such as cotrimoxazole without bacteriological investigation since these infections are so routinely encountered in medical practice (Gupta et al., 2001). Urinary tract includes the organs that collect and store urine and release it from the body which include: kidneys, ureters, bladder and urethra. UTIs are among the most common bacterial and some parasitic infections in humans, both in the rural community and hospital settings and had been reported in all age groups in both sexes (Hooton and Stamm, 1996).

1.2         Statement of the Research Problem

Urinary tract infections in childhood are common and may be difficult to diagnose in young children because of non-specific symptoms. Symptoms such as fever, vomiting, screaming, anorexia and irritability may indicate a urinary tract infection, but they are also common in other childhood diseases. Urinary tract infections in children are significant source of morbidity, particularly when associated with abnormalities. The bacterium Escherichia coli which is part of the normal flora of the body is responsible for most of the reported urinary tract infection especially in primary school children and or appropriate school age children. Escherichia coli is

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normally harmless in the small intestine where it normally resides and becomes a problem when it gains spread to the urinary tract (Shaw et al., 1998).

Drug resistance is a large and growing problem with urinary tract infection. The prevalent pathogens of UTIs have been found to be resistant to chemotherapeutic agents (Okonko et al., 2009), though the antimicrobial susceptibilities of these pathogens are highly predictable. Development of resistance to these antimicrobial agents in UTI cases will therefore affect the future treatment and management of the infections with drugs. Adequate treatment and control of these conditions need good knowledge of the organisms, their epidemiological characteristics and their antibacterial susceptibility testing is therefore mandatory (Singh et al., 2009).

1.3         Justification of the Research

Children are particularly vulnerable to schistosomiasis because of their habit of playing in water, where they may contract the infection. As such, they are the ideal target group to investigate the prevalence of schistosomiasis, and the data collected from this age group can be used to assess not only whether schistosomiasis threatens the health of schoolchildren, but can also be used as a reference for evaluating the need for community intervention (Nokes et al., 1992; Engels et al., 2002: Stothard et al., 2009). There is therefore an urgent need for a renewed commitment to control schistosomiasis and concurrent bacteriuria among children in the country. Parasitic worm infection and bacteria are common in tropical and subtropical countries and many people suffer from multiple parasite infections concurrently. It is important to gain knowledge on co-endemicity patterns because such information can help to design and prioritize interventions and control strategies focusing on areas at highest risk of co-infection. With the pressing need for systematic approaches to health systems, it is necessary to first identify the scope of health

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