COMPARISON OF THE IN/VITRO SENSITIVITY TESTING OF AMINOGLYCOSIDES AGAINST FLUOROQUINOLONE AGENTS IN URINARY TRACT INFECTION ISOLATES

ABSTRACT

In this study a total number of 70 samples were collected from different individuals while 34(48.5%) urine samples were from males, 36(51.4%) were females. 32(45.7%) were found positive for urinary tract infections. The most prevalent isolate was Escherichia coli (75%), followed by Proteus mirabilis (12.5%), and the least isolates were Klebsiella pneumoniae (6.2%) and Staphylococcus saprophyticus(6.2%). The urine samples were cultured using cysteine lactose electrolyte deficient medium (CLED), Blood agar and MacConkey agar. Biochemical tests and gram staining were done to identify the isolates. Antimicrobial sensitivity testing was carried out by disc diffusion technique using antibiotic agents such as aminoglycosides (Gentamicin and Streptomycin) and fluoroquinolones (Ofloxacin and Ciprofloxacin) repectively, to determine the invitro sensitivity pattern of the isolates. The results obtained shows that among the isolates tested, maximum susceptibility was found for fluoroquinolone agents than those of aminoglycosides.

TABLE OF CONTENTS                                                                                         PAGES

Title page    i     

Certification    ii

Dedication    iii

Acknowledgement    iv

Table contents     v

List of table                                                                                                                   viii

List of figures                                                                                                                 ix

Abstract       x

CHAPTER ONE

1.0 Introduction      1

1.1 Aim and Objectives      4

CHAPTER TWO

2.0 Literature Review     5

2.1 UTI in Pregnancy                                                                                               8

2.2 Hospital Acquired UTI                                                                                      10

2.3 UTI in Geriatric Patient                                                                                     11

2.4 Isolates                                                                                                               11

2.5 Antimicrobial resistance among UTI pathogens                                               12

2.6 UTI Therapy                                                                                                      14

2.6.1 Fluoroquinolones                                                                                               14

2.6.1.1 General Structure    15

2.6.1.2 Drug-Drug Resistance                                                                                       15

2.6.1.3 Mechanism of Resistance    16

2.6.1.4 Effects of Quinolones on Bacteria                                                                     16

2.6.1.5 Use of Quinolone in the Treatment of UTI                                                       17

2.6.2 Aminoglycosides                                                                                               18

2.6.2.1 Mechanism of action                                                                                         18

2.6.2.2 Route of Administration                                                                                    19

2.6.2.3 Use of Aminoglycosides in the Treatment UTI                                                19

CHAPTER THREE

3.0 Materials and Methods                                                                                       21

3.1 Study Area                                                                                                          21                                                                      

3.2 Sample Collection                                                                                               21

3.3 Sterilization of Media and Materials      21

3.4 Inoculation of Urine Sample and Isolation of Bacterial Isolates      22

3.5 Microscopy      22

3.6 Identification and Characterization of Isolates                                    23

3.7 Antibiotic Sensitivity Testing                                    23

3.8 Biochemical Test      23

3.8.1 Catalase Tests                                23

3.8.2 Coagulase Test                                                                                                     24

3.8.3 Citrate Test                                      24

3.8.4 Indole Test       25

3.8.5 Methyl red Test          25

3.8.6 Voges – Proskauer Test            25

CHAPTER FOUR

4.0 Results        26

CHAPTER FIVE

5.0 Discussion                          34

5.1 Recommendation            36

5.2 Conclusion        37

References                                                                                      38

LIST OF TABLES

Table Title              Page

1. Age and Sex distribution of study participants --------------------------------------------    28

2. Urinary tract (UTI) Isolates ------------------------------------------------------------------------29

3. Biochemical characterization    ----------------------------------------------------------------    30

4. Colonial morphology of bacterial Isolates    ------------------------------------------------    31

5. Antibiotic sensitivity testing    ------------------------------------------------------------------    32

LIST OF FIGURES

Figure         Title                   Page

1. Percentage distribution using histogram to compare the invitro

Sensitivity testing of Aminoglycoside againstfluoroquinolone agents

In urinary tract Isolates        ----------------------------------------------------------------------33

CHAPTER ONE

1.0 INTRODUCTION

Urinary tract infections (UTI) are conditions where one or more structures in the urinary become infected after bacteria overcome its natural strong defences and it is also a bacterial inflammation in the urinary tract (Bryan, 2011). Urinary tract infection has become the leading infection and major cause of mobility in convalescent care facilities and nursing homes. Furthermore, because antibiotics therapy is given so frequently for symptomatic bacteriuria, the emergence of resistant organisms is becoming increasingly accepted as a consequence of indwelling bladder catheters in an elderly population. Urinary tract infections are the most common of all infections and can occur at any time in the life of an individual. Almost 95% of cases of urinary tract infection are caused by bacteria which typically multiply at the opening of the urethra and travel up to the bladder. Urinary tract infections are more than 100 organisms per milliliter of urine in a symptomatic patient or less than 100,000 organisms per milliliter of urine in asymptomatic patient (Bryan, 2011).

Patient’s symptoms include urgent, painful, and frequent urination along with malodorous and/or cloudy urine; signs of infection include the presence of blood (hematuria) or white blood cells (pyuria) in urine (Jacobsen et al., 2008).Urinary tract infections comprise a spectrum of diseases of varying severity with different outcomes and treatment guidelines. Asymptomatic infections are referred to as asymptomatic bacteriuria (ABU) where as symptomatic infections are classified as either cystitis if they are confined to the bladder or pyelonephritis if the infection has spread to the kidney. Due to the absence of symptoms asymptomatic bacteriuria is often only discovered through a positive urine culture and does not require treatment unless risk factors for complication are present (example pregnancy and kidney transplantation). (Foxmanet al., 2003).

Any situation in which the urine does not naturally flow increases the chance of infection in this circumstance, urine accumulates and distends the elastic bladder, even a few bacteria can multiply to high levels, during this time causing infection. Urine provides abundant nutrient for any specie of bacteria. Bacteria are the most common cause of urinary tract infections; the most common is bladder infection, cystitis. Urinary tract infections can also involve the urethra and kidney, these infection are more difficult to treat, most urinary tract infection occur mostly in females than male because the design of the female urinary tract makes it easier for bacteria to enter and cause an infection. It is estimated that about 5% - 6% of all sexually active women have bacteriuria between ages 20 and 40. (Bryan, 2011).

Bacteria are the most common cause of urinary tract infection. The most common bacterium that causes urinary tract infection is Escherichia coli (E. coli), other bacteria that can cause this infection include: Proteus spp, Klebsiellaspp, Staph. aureus, Staph. saprophyticus. The gram negative bacteria Escherichia coli are responsible for between 80% - 85% of cystitis cases. Urinary tract infections account for a large proportion of antibacterial drug consumption and have large socio-economic impacts. Anatomical factors can cause urinary tract infection, any anatomical problem that obstructs the flow of urine can make it easier for bacteria to become trapped leading to a urinary tract infection. An uncircumcised penis can trap bacteria leading to urinary tract infection.Antimicrobial agents can be grouped by their mode of action, their ability to inhibit the synthesis of the cell wall, cell membrane protein and nucleic acid of bacteria. Antibiotics are commonly prescribed for a variety of diseases, basically in the case of urinary tract infection the antimicrobial agents; aminoglycosides (gentamicin, neomycin, streptomycin) and fluoroquinolones (ofloxacin, ciprofloxacin) are routinely used for the treatment of urinary tract infections. Antimicrobial agents are generally regarded as bacteriostatic and bacteriocidal agents. Bacteriostatic agents inhibits the growth and multiplication of susceptible microorganisms, example include; chloramphenicol, erythromycin, sulphonomide and tetracyclines. Bacteriocidal agents kill and inhibit microorganisms. Example include; cephalosporins, penicillins, polymyxins. For example, cases of urinary tract infection are reportedly said to occur more in convalescent care facilities. Furthermore aerobic gram negative rods resistant to first generation cephalosporin, trimethoprim-sulfamethoxazole, and ampicillin constitute a major proportion of the etiologic agents. These organisms include Enterobacterspecies, pseudomonas aeruginosa, providenciastuartii, morganellamorganni, and proteusspecies. Thus, oral antimicrobial agents have become notably less useful, so that parenteral agents are often required for patients with symptomatic bacteriuriaand systemic signs of infection(Jacobsen et al., 2008).

Aminoglycoside agents have been used increasingly in the treatment of urinary tract infection because organisms are resistant not only to the standard oral agents but often to parenteral beta-lactam agents as well. Furthermore the aminoglycosides are considerably more toxic than the beta-lactam agents. This toxicity is magnified in the elderly patient in whom renal dysfunction and hearing problems may occur as a natural consequence of aging.(Hiltet al., 2013)

Fluoroquinolone agents have a broad antibacterial spectrum that includes activity against many aerobic gram-negative rods, including those resistant to aminoglycosides and cephalosporin agents. (Hiltet al., 2013). And the basis of this project work is to compare the invitro efficacy of aminoglycoside and fluoroquinolone agents in the isolates of urinary tract infections.

1.1 AIM AND OBJECTIVE

Aim: The aim of this work is to evaluate the invitro sensitivity pattern of aminoglycosides against fluoroquinolone agents on UTI isolates

Objectives

1. To isolate and identify bacteria pathogens causing UTI.

2. To determine the prevalence of various bacteria in urinary tract infection

3. To determine the invitro sensitivity pattern of the isolates with fluoroquinolones and aminoglycosides.

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