ANTIBIOTIC SUSCEPTIBILITY PROFILE OF BACTERIAL ISOLATES FROM URINE SPECIMENS OF PATIENTS

ABSTRACT

This study was conducted to evaluate the major etiologic agent of urinary tract infections amongst patients that attend Michael Okpara University of Agriculture Umudike Medical clinic. About 100 samples of early morning mid-stream urine samples of about 10-15 ml were collected; using UV sterilized plastic bottles with air-tight screw cap tops from Michael Okpara University of Agriculture Umudike Health Center. All samples collected were transported to the laboratory immediately and cultured using the streak plate method on Cysteine Lactose Electrolyte Deficient agar and blood agar and incubated for 24 hours at 37ᵒC for bacterial growth. The resulting colonies from the Cysteine Lactose Electrolyte Deficient agar and blood agar were purified by sub-culturing on freshly prepared nutrient agar plates. The plates were incubated at 35°C for 24hours. It was observed that that Escherichia coli is the most frequently occurring isolates with a percentage occurrence of 15(27.3%), followed by Pseudomonas sp with a percentage occurrence of 12(21.8%), then Staphylococcus aureus 10(18.2%), Klebsiella sp 9(16.4%) and Proteus sp 7(12.7%), whereas Streptococcus sp has the least percentage occurrence of 2(3.6%). The antibiotic sensitivity test reviewed that Gentamycin (81.8%) is the drug of choice for the treatment of skin infection following to the fact that it show high rate of sensitivity to the bacterial isolates tested, this is followed by Lefofloxascin (78.2%), Ciprofloxacin (72.7%), Cefoxitin (50.9%), Cefotaxime (47.3%) Azithromycin and Ceftriaxone (40%), Cefuroxime (32.7%), Cefexime (30.9%) where as Imipenem (23.6%) has the least percentage susceptibility to the isolates. the isolates however were moderately resistant to Imipenem (32.7%), Cefuroxime (29.1%), Cefotaxime (29.1%), Azithromycin (27.3%), Cefexime (25.5%) while few showed low resistance to Lefofloxascin (3.6%), Ciprofloxacin (9.1%) and Cefexime (12.7%). There should be reinforcements for mandatory urine culture for all suspected Urinary tract infections to properly guide the therapy. Further studies on urinary tract infections should be encouraged to properly discover more etiological agents responsible for urinary tract infection among patients under study.

TABLE OF CONTENTS

Title Page                                                                                                                                 i

Certification                                                                                                                           ii

Dedication                                                                                                                              iii

Acknowledgements                                                                                                                iv

Table of Contents                                                                                                                   v

List of Tables                                                                                                                          vii

List of Figures                                                                                                                         viii

Abstract                                                                                                                                  ix

CHAPTER ONE

1.0       Introduction                                                                                                                1

1.1       Aim and Objectives                                                                                                    4

CHAPTER TWO

2.0       Literature Review                                                                                                       5

2.1       Antibiotic Sensitivity of Pathogens Causing Urinary Tract Infection                     5         

2.2       Isolation and Identification of Uropathogens                                                             6

2.3       Susceptibility Pattern of Uropathogens to Ciprofloxacin                                           9

2.4       Bacterial Pathogens in Urinary Tract Infection and Antibiotic Susceptibility

Pattern                                                                                                                         10

2.5       Antibiotic Susceptibility Test                                                                                     12

CHAPTER THREE

3.0       Materials and Methods                                                                                               14

3.1       Collection of Samples                                                                                                14

3.2       Sterilization of Materials                                                                                            14

3.3       Preparation of Culture Media                                                                                     14

3.4       Inoculation and Isolation                                                                                            15

3.5       Purification of Isolates                                                                                                15

3.6       Identification of the Isolates                                                                                       15

3.7       Gram Staining                                                                                                            15

3.8       Biochemical Test                                                                                                        16

3.8.1    Catalase Test                                                                                                               16

3.8.2    Indole Test                                                                                                                  16

3.8.3    Citrate Utilization Test                                                                                               16

3.8.4    Hydrogen Sulphide (H₂S) Production Test                                                                17

3.8.5    Starch Hydrolysis                                                                                                       17

3.8.6    Motility, Indole, Urease (MIU)                                                                                  17

3.8.7    Coagulase Test                                                                                                           18

3.8.8    Oxidase Test                                                                                                               18

3.9       Antibiotic Susceptibility Testing                                                                                18

CHAPTER FOUR

4.0       Results                                                                                                                        20

CHAPTER FIVE

5.0       Discussion, Conclusion and Recommendation                                                          29

5.1       Discussion                                                                                                                   29

5.2       Conclusion                                                                                                                  31

5.3       Recommendations                                                                                                      31

            References                                                                                                                  32

            Appendix                                                                                                                    39

LIST OF TABLES

LIST OF FIGURES

CHAPTER ONE

1.0       INTRODUCTION

Urinary tract infection (UTI) represents one of the most common diseases encountered in medical practice. This diseases affects people all age groups, being most common in women (Khan et al., 2014). The most common pathogens causing UTI are E. coli and other enterobacteriacae, which accounts for approximately 75% of the isolates (Beyene and Tsegaye, 2011). Other gram negative organisms like P. aeruginosa and gram positive organisms like Enterococcus spp. are the common organisms causing UTI in hospital settings (Minardi et al., 2011). In most cases of UTI, empirical antimicrobial therapy is initiated before culture and sensitivity results are available. Poor patient compliance and incomplete course of antibiotic therapy have resulted in the evolution of increase in resistance to urinary isolates (Singhal et al., 2014).

For uncomplicated lower UTI, such as acute cystitis in otherwise healthy women, not only a revival of old (oral) antibiotics, such as fosfomycin trometamol, pivmecillinam, nitrofurantoin, can be observed in many guidelines, but even a non-antimicrobial measure has been tested in a pilot study. It will therefore be interesting to see the results of forthcoming phase III studies and whether antibiotic therapy could at least be partially replaced. For prophylaxis of recurrent episodes of uncomplicated UTI, non antimicrobial measures are already preferred and antimicrobial prophylaxis is only recommended as a last resort. However, for complicated, nosocomial and severe UTI including pyelonephritis, antibiotic therapy will still be a corner stone in combination with treatment of the underlying complicating conditions. Unfortunately, there are few new antimicrobial drugs in the pipelines of pharmaceutical companies with prospects to overcome the problem of multi and extended drug resistant uropathogens (Hamdan et al., 2011). Although the classical distinction between uncomplicated and complicated UTI is still valid in principle, the different criteria to be considered are so heterogeneous, that a better (phenotypical) sub-classification might be helpful, as proposed by the European Section of Infection of Urology (ESIU) of the European Association of Urology (EAU). In consideration of so many new aspects related to optimal management of UTI, it has been our pleasure to edit a joint presentation of the results from different research groups in one special scientific publication challenging established as well as new scientific approaches to improve prophylaxis and treatment of patients suffering from UTI.

Microorganisms form a bulk of the earth’s biomass and their ability to adapt to newly found environments makes them beneficial or pathogenic. Many human diseases are as a result of infections caused by bacteria pathogens, either external or internal of the human host. One of such bacterial infection is the Urinary Tract Infection (UTI), involving the presence of bacteria in the urinary tract (UT), which is naturally sterile.UTI mostly occurs in patients with anatomically and functionally normal UT and usually results from spontaneous ascent of bacteria from the urethra to the bladder. Occasionally, the bacteria progresses to the kidney and bloodstream. However, since asymptomatic colonization of the UT can occur, other features such as the presence of inflammatory markers or follow-up cultures are needed to correctly diagnose a person with UTI. Bacteria colonization of the UT is predominantly caused by Gram-negative species, such as Escherichia coli, Klebsiella, Proteus and Pseudomonas and rarely, by Gram-positive organisms such as haemolytic Streptococci and Staphylococcus saprophyticus. Global records on the disease show that among children, the infection is more common in young girls, except in the neonatal age group where boys predominate. It is also estimated that about 20% of women develop an UTI during their lifetime; the incidence increases at puberty and remains high throughout adult life. Furthermore, UTIs account for approximately 23% of all hospital-acquired infections. In Ghana, 7.3% of pregnant women attending antenatal care have been identified to have significant bacteriuria (presence of bacteria in urine). E. coli as dominant bacteria isolate, account for about 37% of all cases. Infants, pregnant women, patients with spinal cord injuries, diabetes, multiple sclerosis, acquired immunodeficiency disease syndrome or underlying urologic abnormalities are subjects that are at increased risk for UTI. In addition, catheter-associated UTI is the most common nosocomial infection. Many previous studies have shown that E. coli is the most common etiological agent of UTI in both hospital and community acquired infections. Hospital acquired UTI has also been characteristically associated with a higher prevalence of enterococci and Coagulase- Negative Staphylococci. In addition, Klebsiella pneumonia, Streptococcus agalactiae, Proteus mirabilis, viridians streptococci, Klebsiella oxytoca, Pseudomonas aeruginosa, Citrobacter freundii, Enterobacter cloacae, and Staphylococcus aureus have been identified as etiologic agents of UTI. Due to the rapidly evolving adaptive strategies of bacteria, the etiology of UTI and antibiotic resistance profile of bacterial uropathogens have changed considerably over the past years, both in community and nosocomial infections. Many studies conducted from the USA and Europe have revealed increasing antibiotic resistance among uropathogenic E. coli to ampicillin, trimethoprim, and sulfonamides.

Apparent shift in the etiological agents of urinary tract infection and associated problem of antibiotic resistance amongst bacterial uropathogens from time to time and from one institution to another have initiated health institution to carry out continuous evaluation of UTI from the view point of their spectrum and drug susceptibility testing. Accurate identification of bacterial uropathogens and determining their drug susceptibility pattern are critical for efficient management of patients with UTI. They are also associated with significant clinical and financial benefits, via the reduction of mortality rates and overall hospitalization costs (Davoodian et al., 2012). In view of this, identification and antimicrobial susceptibility testing of clinical isolates by means of fully automated systems have become a common practice in many laboratories. The VITEK 2 compact system is a new automated system designed to provide accurate identification and susceptibility testing results for most clinical isolates of both Gram-positive and Gram-negative bacteria. Apart from accurate identification and susceptibility testing shortened turnaround times, improved specimen handling, enhanced quality control, reproducibility and the ability to track results are further advantages of the system.

1.1       AIM AND OBJECTIVES

To determine the major etiologic agent of urinary tract infections amongst patients that attends Michael Okpara University of Agriculture Umudike Health Center.

The specific objectives are;

1.     To isolate and identify bacterial causing urinary tract infection.

2.     To determine the percentage occurrence of the isolates.

3.     To ascertain the antibiotic sensitivity and resistance pattern of the isolates,

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