BACTERIA ASSOCIATED WITH URINARY TRACT INFECTION AMONG MALE STUDENTS IN MOUAU

ABSTRACT

This study was carried out to evaluate the types and prevalence of bacteria causing Urinary tract infection(UTI) among male students in the University. A total of 65 randomly selected  male students of Michael Okpara University who participated in this project work. Diagnosis of UTI was made by urinalysis and urine culture. Identification of the isolates was carried out by microscopy and culture including biochemical tests. Antibiotic susceptibility tests were done by disc diffusion technique. In Gram negative bacteria, the predominant isolate was the Escherichia coli (40.9%) followed by  Klebsiella spp. (9.1%) and Proteus spp. (9.1%)  In Gram positive bacteria the main organism identified was Staphylococcus aureus (36.4%) followed by Streptococcus spp. (4.5%). Escherichia coli is the most common cause of UTIs. Among the tested antibiotics, the highest susceptibility for the bacteria was shown by Gentamicin and Ciprofloxacin while the highest resistance for the bacteria was shown by Augmentin, Nalidixic acid, Trimethoprim-sulphamethoxazole (Septrin), Ampicillin and Amoxicillin. In order to decrease resistance to antibiotics, the use of antibiotics should be kept under supervision and should be given in appropriate doses for an appropriate period of time.

TABLE OF CONTENTS

Title Page.                                                                                                                   i

Certification.                                                                                                                        ii

Dedication.                                                                                                                       iii

Acknowledgements.                                                                                                               iv

Table of Contents.                                                                                                                 v

List of Tables.                                                                                                                         vi

Abstract.

CHAPTER ONE: INTRODUCTION.                                                                                                         

1.1 Introduction.                                                                                                                    1

1.2 Aim and Objectives.                                                                                                      4

1.3 Research Hypothesis.                                                                                                       5

CHAPTER TWO:  LITERATURE REVIEW.                                                                                               

2.1 Literature review.                                                                                                              6

2.2 Epidemiology of UTI.                                                                                                       9

2.3 Pathogenesis of UTI.                                                                                                               10

2.4 Causes of UTI.                                                                                                                          11

CHAPTER THREE:  MATERIALS AND METHODS.                                                                               

3.1 Collection of Samples.                                                                                                           13

3.2 Microscopic Examination.                                                                                                    13

3.3 Culture.                                                                                                                                        13

3.4 Gram Staining.                                                                                                                          14

3.5 Biochemical testing.                                                                                                                14

3.5.1 Catalase test.                                                                                                                          15

3.5.2 Coagulase test.                                                                                                                       15

3.5.3 Oxidase test.                                                                                                                           16

3.5.4 TSI test.                                                                                                                                   16

3.5.5 SIM (Hydrogen sulphide,Indole and Motility) test.                                                  16

3.5.6 Citrate test.                                                                                                                              17

3.5.7 Urease test.                                                                                                                             17

3.6 Antimicrobial Susceptibility Testing.                                                                                18

CHAPTER FOUR: RESULTS                                                                                                   19

CHAPTER FIVE :DISCUSSIONS, CONCLUSION AND RECOMMENDATION.                        

5.1 Discussion.                                                                                                                                 25

5.2 Conclusion.                                                                                                                                27

5.3 Recommendation.                                                                                                                    28

REFERENCES.

LIST OF TABLES

Table 1: Age distribution of participants in the study.

Table 2:Distributions of uropathogens among the positive isolates.

Table 3:Distribution of Gram positive and Gram negetive bacteria among the isolated Uropathogens

Table 4: In-vitro Antibiotics Sensitivity pattern of  bacteria isolates in the study.

Table 5:Result comparison between students living Off-Campus to those in the hostel.            

APPENDIX

CHAPTER ONE

 INTRODUCTION

Urinary Tract Infection (UTI) remains the most common bacterial infection in human population and is also one of the most frequently occurring nosocomial infections (Gastmeir et al.,1998). Its annual global incidence is of almost 250 million (Ronald et al.,2001). UTIs refer to the presence of microbial pathogens within the urinary tract and it is usually classified by the site of infection as bladder (cystitis), kidney (pyelonephritis) or urine (bacteriuria). They are asymptomatic or symptomatic. UTIs that occur in a normal genitourinary tract with no prior instrumentation are considered as “uncomplicated”, whereas “complicated” infections are diagnosed in genitourinary tracts that have structural or functional abnormalities including instrumentation such as indwelling urethral catheters, and are frequently asymptomatic (Gonzalez et al.,1999). Many different microorganisms can cause UTIs though the most common pathogens causing the simple ones in the community are Escherichia coli and other Enterobacteriacae, which accounts for approximately 75% of the isolates (Getenet et al.,2011). Treatment of UTI is often started empirically and therapy is based on information determined from the antimicrobial resistance pattern of the urinary pathogens (Wilson et al.,2004). The prevalence of antimicrobial resistance among urinary pathogens has been increasing worldwide due to aberrant use of antibiotics in practice (Bonadio et al.,2001).Distribution of urinary pathogens and their susceptibility to antibiotics varies regionally so it becomes necessary to have knowledge of distribution of these pathogens and their susceptibility to antibiotics in a particular setting(Farrell et al.,2003).

Urinary tract infection (UTI) is one of the commonest infections encountered by clinicians and despite the widespread availability of antimicrobial agents UTI has become difficult to treat because of appearance of pathogens with increasing resistance to antimicrobial agents.

In many samples, millions or billions of bacteria are present. When plated, even a small sample would give rise to so many colonies that they would run together and be hard to count. What is usually done is to plate a series of dilutions of the sample. You can always take the dilution factor into account when calculating the number of cells in the original sample.  By doing a series of dilutions, you insure that one of the dilutions will give you colony numbers that are not too high or too low, but just right.

The results are reported in cells/mL. For example, if 0.001 mL (1 microliter, abbreviated as 1 μl) is plated, the colony number must be multiplied by 1000, since 0.001 mL is 1/1000th of a mL. This serial dilution technique is used when we have no idea how many bacterial cells are present. Plating serial dilutions increases the chance of (1) quantitating the bacteria and (2) isolating colonies of the different types of bacteria present.

In the clinical laboratory, this principle is applied to the microbial assay of urine. A urine sample is streaked on medium using a special inoculating loop known as a “calibrated loop.” The calibrated loop is calibrated so that it picks up only 0.001 mL of urine from the patient’s specimen. Thus, while no actual dilution is done, only a small volume (one microliter) is streaked onto a plate. When reading the results of the urine culture, the medical technologist takes into account that only 0.001 mL was plated, multiplies the results by 1000 and reports “colonies/mL.”

Although we would expect fluid stored in the bladder to be sterile, it must pass through the urethra as it exits the body. The distal portion of the urethra is colonized by normal flora. As the urine passes through this region, some of the normal flora gets caught up in the flow. Therefore, we would expect to find some bacteria in urine. The medical microbiologist must determine if the bacteria in a urine sample indicate a urinary tract infection (UTI). Complicating matters is that most of the bacteria that cause UTIs are part of the normal flora of the intestinal tract, likely the same ones that normally inhabit the distal portion of the urethra.

In the clinical lab, 0.001-mL urine samples are streaked on the media and incubated at 37° C for 24 hours. At the end of the incubation period, the medical technologist counts the number of colonies on the plate. (Each colony represents 1000 cells present in the original specimen) If the technologist counts 30 colonies on the plate, there were 30 cells in the 0.001 mL plated. This 0.001 mL is one-thousandth of a mL, so to calculate the number of cells per mL of the original urine sample, the technologist multiplies 30 colonies in 1/1000th mL x 1000 = 30,000 cells per mL of the original urine sample. The threshold (the minimum number of cells that must be present in order to suspect a UTI) is 100,000 cells/mL. Thus, a urine culture plate with more than 100 colonies on it represents an original urine sample with more than 100,000 bacterial cells per ml. This far exceeds the number that can be picked up from the distal urethra. The bacteria are assumed to have come from the bladder and represent the organism causing the UTI.

 1.1  AIMS AND OBJECTIVES:

The aim of this study is to determine bacterial etiologic agents responsible for urinary tract infection among male student in MOUAU and to evaluate their in vitro susceptibility pattern to commonly used antimicrobial agents.

Objectives;

·       To isolate and identify bacteria pathogens in UTI.

·       To determine the prevalence of various isolates.

·       To determine the antibiotic resistance pattern of the isolates.

1.2  RESEARCH  HYPOTHESIS:

·       Enterobacteriaceae are the most common cause of Urinary tract infection.

·       E.coli is the most prevalent uropathogen isolated from the Urinary tract.

·       Most Uropathogen are resistant to the commonly used antibiotics due to misuse of these drugs.

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