BIOCHEMICAL CHARACTERIZATION OF BACTERIA ISOLATED FROM URINE SPECIMENS OF FEMALE STUDENTS IN MICHAEL OKPARA UNIVERSITY OF AGRICULTURE UMUDIKE, ABIA STATE

ABSTRACT

This project work was carried out to study the biochemical characterization of bacteria isolated from urine samples of female students in Michael Okpara University of Agriculture, Umudike, Abia State. A total of 110 samples were collected randomly from the female students. The specimens were cultured on Cysteine lactose electrolyte deficient agar. Result from this study indicated that 58(52.7%) of the specimens were significant for Urinary Tract Infections (UTIs) and 52 (47.3%) were not significant for UTI. Result from biochemical characterization of the Gram positive bacteria showed that 36(75%) showed positive for catalase while 12(25%) showed negative for catalase tests. Coagulase test showed that about 17(35.4%) of the isolate were positive for coagulase while 31(64.6%) tested for coagulase were negative. Citrate utilization test was carried out on 16 Gram negative bacteria. The result revealed that 9(56.3%) of the isolate was positive with only 7(43.7%) showing negative result. Indole test for 16 isolates showed positive for 12(75%) of the total isolate. While, 4(25%) were negative. Result for the hydrogen sulphide (H2S) showed that no presence of hydrogen sulphide was observed in any of the specimens after the biochemical test. Staphylococcus aureus, Coagulase negative Staphylococcus (Staphylococcus saprophyticus), Escherichia coli, Enterococcus species, Klebsiella pneumonia, Pseudomonas aeruginosa, Citrobacter Koseri and Group B Streptococci were the various bacteria isolated from the specimens. This study revealed a high prevalence rate of UTI among the tested female students most of which was caused by Staphylococcus saprophyticus.

TABLE OF CONTENTS

Title page i

Certification ii

Dedication iii

Acknowledgements iv

Table of contents v

List of tables vii

Abstract viii

CHAPTER ONE  

INTRODUCTION

1.0 Introduction 1

1.1 Aims of Study 3

1.2 Objective 4

CHAPTER TWO

LITERATURE REVIEW

2.1 Urinary Tract Infection 5

2.2 Epidemiology of the Urinary Tract System 7

2.3 Causative Agents of Urinary Tract Infections 8

2.4 Route of Infection 9

2.5 Symptoms of Urinary Tract Infections 10

2.6 Risk Factors of Urinary Tract Infection 12

2.7 Diagnosis of Urinary Tract Infection 14

2.8 Types of Urinary Tract Infections 15

CHAPTER THREE

MATERIALS AND METHODS

3.1 The Study Site 17

3.2 The Study Population 17

3.3 Specimen Collection 17

3.4 Sterilization of Materials and Preparation of Media 18

3.5 Culturing of Urine Specimen and Isolation of Bacterial Isolates 18

3.6 Gram Stain of Sub-Cultured Isolates 18

3.7 Biochemical Characterization and Identification of Bacterial Isolates 19

3.7.1 Catalase Test 19

3.7.2 Coagulase Test 19

3.7.3 Citrate   Test 20

3.7.4 Indole Test 20

3.7.5 Hydrogen Sulphide test 20

CHAPTER FOUR

RESULTS 22

CHAPTER FIVE

5.1 Discussions 29

5.2 Conclusions 30

5.3 Recommendations 31

     References

LIST OF TABLES

TABLE   TITLE             PAGE

    1 Gram staining reaction and biochemical characterization

 of the isolate. 23

    2  Frequency and percentages of the species of isolates among female

 Students with Asymptomatic Urinary Tract Infection 27

CHAPTER ONE

INTRODUCTION

A Urinary Tract Infection is defined as a microbial infiltration of the urinary tract and is one of the most common bacterial infections of the urethra (Urethritis), bladder (Cystitis), Ureter (Ureteritis), kidney (Pyelonephritis) (Barber, et al., 2013). The two most common symptoms of lower Urinary tract infections or bladder infections are burning with urination and frequent urination in the absence of vaginal discharges and significant pain. Symptoms of the upper urinary tract infection include; fever, flank pain, nausea or vomiting, bloody urine and visible pus in the urine. If a urine contain significant bacteria but there are no symptoms, the condition is known as an asymptomatic bacteriuria, but if there are symptoms, it is known as a symptomatic bacteriuria. The frequency and natural history of asymptomatic bacteriuria vary from different populations (Ezeadila, et al., 2015). Bacteria isolated from the urine of individuals with asymptomatic bacteriuria usually originate as colonizing flora of the gut, vagina or periurethral area. Bacteria that contaminate urologic fluids may be introduced into the genitourinary tract for patients that are subjected to urinary tract instrumentation. Without prior host colonization, the genitourinary tract is always sterile, apart from the distal urethra. Asymptomatic bacteriuria occurs following ascension to the kidneys. Microorganisms then persist in the urinary tract without eliciting a host response sufficient to produce symptoms or cause an irritation. Variables such as; incomplete bladder emptying or the presence of a foreign body, host genetic predisposition may facilitate persistent urinary tract infections (Ezeadila, et al., 2015). Urinary tract infections are one of the major causes of illness affecting all groups. It is more common in females than males at a ratio of 8:1. Urinary tract infections are reported more in women and could be due to the proximity of the genital tract and urethra-anus or the anatomical predisposition or urothelial mucosa adherence to muco-polysaccharides lining or other host factors. Urinary tract infections may involve only the lower urinary tract or both the upper and lower tract (Angoti, et al., 2016). As a woman’s estrogen level decreases with menopause, her risk of UTI increases due to the loss of the protective vaginal flora (Angoti, et al., 2016; Ezeadila, et al., 2015). They occur most frequently between the ages of 16-34 years with 10% of women getting an infection yearly and about 50-60% report at least one UTI in their lifetime and one in three will have at least one symptomatic UTI necessitating antibiotic treatment by age 24 (Al-badr and Al-shaikh, 2013). Normally, the urinary tract is sterile, but bacteria may rise from the perianal region, possibly leading to urinary tract infections. Pathogens in the bladder may stay silent or can cause irritative symptoms like; urinary frequency and urgency, and 8% of women may have asymptomatic bacteriuria. If a bacterium enters the blood stream, it could cause severe complications such as; shock, septicaemia and rarely death. Patients with catheter or those suffering from complaints of prostatitis are also prone to urinary tract infection. Transmission occurs through four ways namely: sexual intercourse, from mother to the foetus by placenta, through poor personal hygiene and by communal sponge and towel usage (Priyadharsini, et al., 2014). Risk factors for developing a catheter associated urinary tract include; prolonged catherization, female gender, older age and diabetes. Urinary tract infections (UTIs) are caused by both Gram positive and Gram negative bacteria, as well as by certain fungi, with a large proportion caused by Enterobacteriaceae. The most common causative agent for both complicated and uncomplicated UTIs is the Uropathogenic Escherichia coli (UPEC). For the agents involved in uncomplicated UTIs, UPEC is followed in prevalence by Klebsiella pneumonia, Staphylococcus aureus, Staphylococcus saprophyticus, Enterococcus faecalis, Group B Streptococci, Proteus mirabilis, and Pseudomonas aeruginosa. For complicated UTIs, the order of prevalence following UPEC is Enterococcus spp, Klebsiella pneumoniae, Staphylococcus aureus, Proteus mirabilis, Pseudomonas aeruginosa and Group B Streptococci (Flores-Meres, et al., 2015). It is believed that the bacteria are usually able to attach to the bladder wall to form a biofilm that resists the body’s immune response. The mainstay of treatment of urinary tract infections is antibiotics. However, increasing antibiotic resistance is causing concern about the future of treating patients with complicated and recurrent urinary tract infections (Ezeadila, et al., 2015). Among uropathogens, the rate of resistance is high and frequency of resistance to antibiotics and drugs is directly linked to consumption of antibiotics. Although, there is a large group of antimicrobial agents available for the treatment of UTIs, none of them can treat all UTIs (Patil, et al., 2013). In recent years, antibiotic resistance has become a major problem worldwide due to several factors related to the genetic nature of the organisms and selective antimicrobial pressure in animals and humans. Prevalence of the urinary pathogens and their susceptibility reactions to antibiotics differ from place to place with time. It is essential to know the current trends of UTIs in different locations to ease diagnosis and thus establish the suitable antimicrobial agents for such infections in order to facilitate quick recovery, prevent or reduce complications of antimicrobial resistance (Ochada, et al., 2015).

1.1 AIM OF STUDY

The aim of this project work is to study the biochemical characterization of bacteria isolated from urine specimens of female students in Michael Okpara University of Agriculture Umudike, Abia State.

1.2 OBJECTIVES

1. To Isolate bacteria from urine specimens of female students from Michael Okpara University of Agriculture, Umudike.

2. To carry out biochemical tests to identify the bacteria isolated from the urine specimens.

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