PREVALENCE OF GRAM NEGATIVE BACTERIA IN URINE SPECIMENS AND THEIR ANTIBIOTIC SUSCEPTIBILITY PATTERN

ABSTRACT

Urinary tract infection (UTI) is one of the common bacterial infections in man. The changing antimicrobial sensitivity in UTI demands use of appropriate antibiotics. This study was conducted to determine the prevalence and antimicrobial susceptibility of uropathogens. Clean catch midstream urine samples were collected from 200 students of Michael Okpara University Of Agriculture,Umudike. Antimicrobial susceptibility was performed by disc diffusion method. The results revealed that out of 200 urine samples, 127(63.5%) were found positive. The prevalence was higher in females than in males (females: 44.5; males: 19%). Age group of 21-25 years showed higher prevalence of UTI. The most common organisms isolated were Escherichia coli, Klebsiella pneumoniae , Proteus spp and Pseudomonas aeruginosa. These represented 52.76%, 19.69%, 15.75% and 11.81% of isolates respectively. Ofloxacin, Cefexime, Levofloxacin and Ceftriaxone Sulbactarm were found the most susceptible drug against isolated uropathogens. In conclusion, the gram negative organisms isolated showed to be very sensitive to the following drugs; Ofloxacin, Cefexime, Levofloxacin and Ceftriaxone Sulbactarm and it could therefore be suggested that in the situation of clear UTI symptoms and in the absence of physician that these drugs (Ofloxacin ,Cefexime, Levofloxacin and Ceftriaxone Sulbactarm) could be  used. Therefore isolation of pathogens in urine and subsequent antibiotic testing are vital in the treatment of urinary tract infection.

TABLE OF CONTENTS

Certification                                                                                                                                   i          

Dedications                                                                                                                                  ii         

Acknowledgements                                                                                                                   iii

Table of Contents                                                                                                                      iv

List of Tables                                                                                                                             vii

Lists of Figures                                                                                                                          viii

Abstract                                                                                                                                             ix

CHAPATER ONE
1.0Introduction                                                                                                                                   1         

1.2Aims and Objectives                                                                                                                     3

CHAPTERTWO

2.0Literature Review                                                                                                                          4

2.1Biology Of Urinary Tract                                                                                                              4

2.2Classification Of  UTI                                                                                                                   5

2.2.1Uncomplicated And Complicated Urinary Tract Infection                                                        6

2.2.2 Recurrent Urinary Tract Infection                                                                                             6

2.3 Route Of Infection And Causes                                                                                                    7

2.3.1Intercourse                                                                                                                                  8

2.3.2Sex                                                                                                                                              8

2.3.3Urinary Catheters                                                                                                                       9         

2.3.4Others                                                                                                                                         9

2.4 Hospitalized Conditions That Enhances The Prevalence Of The Infection                                 10

2.4.1Diabetes And UTI                                                                                                                       10

2.4.2Other Factors Associated With Risk of UTI                                                                              11

2.4.3UTI During Pregnancy                                                                                                               12

2.5 Symptoms Of UTI                                                                                                                        13

2.6Microorganisms found in Urine and their Etiology                                                                      14 
2.6.1 Bacteria                                                                                                                                     14

2.6.2viruses                                                                                                                                         15 
2.6.3 Fungi                                                                                                                                          16

2.6.4 Protozoa                                                                                                                                     16

2.7 Entry Of Bacteria Into The Urinary Tract (Pathogenesis )                                                          17

2.7.1 Hematogenous Route                                                                                                                17

2.7.2 Ascending Route                                                                                                                       17

2.8 Diagnosis                                                                                                                                      19

2.8.1 Different Diagnosis Methods                                                                                                    19

2.8.1.1 Diagnosis In Children                                                                                                             20

2.8.1.2 Differential Diagnosis                                                                                                            20

2.9 Treatment                                                                                                                                     21

2.9.1 Asymptomatic Bacteriuria                                                                                                        21

2.9.2Uncomplicated                                                                                                                           22

2.9.3Complicated                                                                                                                               23

2.9.4 Pyelonephritis                                                                                                                            23

2.10 Prevention And Control                                                                                                              23

2.11 Antibiotics                                                                                                                                  24

2.12 Antimicrobial Resistance                                                                                                           24

2.13 Mechanism Of Drug Resistance                                                                                                 25

2.13.1 Efflux Pumps                                                                                                                           25

2.13.2 Modification Of Target Molecule                                                                                           25

2.13.3 Antibiotic Inactivation                                                                                                           25

2.13.3.1β-Lactamases                                                                                                                         26

2.13.3.2 Aminoglycoside Modifying Enzymes (Age's)                                                                     26

2.13.3.3 Chloramphenicol-Acetyl-Transferases                                                                                26

2.13.3.4 Drug Inactivation By Group Transfer                                                                                  26

2.13.3.5 Drug Inactivation By Redox Process                                                                                   27

2.13.4 Reduced Membrane Permeability                                                                                           27

2.15 Control And Prevention Of Emergency And Spreading Of Antimicrobial Resistance                   28

CHAPTERTHREE
3.0 Materials And Methods                                                                                                                30

•       Specimen Collection And Bacterial Isolates                                                                          30       

•       Media                                                                                                                                      30

3.3 Sterilization                                                                                                                                  30

3.4 Preparation Of Culture Media                                                                                                      30

3.5Isolation Of Bacteria From Urine Samples                                                                                   31

3.6 Identification of Isolates                                                                                                               31

3.6.1 Gram Staining                                                                                                                           31

3.6.2 Motility Test                                                                                                                              31

3.6.3 Antimicrobial Susceptibility Test                                                                                              32

3.7 Biochemical Cultural Characteristics                                                                                           32

3.7.1 Catalase Test                                                                                                                              32

3.7.2 Coagulase Test                                                                                                                          32

3.7.3 Citrate Test                                                                                                                                33

3.7.4Indole Test                                                                                                                                  33

3.7.5 Oxidase Test                                                                                                                              33

3.7.6 Urease Test                                                                                                                                33

3.7.7 Methyl Red Test                                                                                                                        34

3.7.8 Voges-Proskaeur Test                                                                                                                34

CHAPTER FOUR

4.0 Results                                                                                                                                          35

CHAPTER FIVE

5.0 Discussion, Conclusion And Recommendations                                                                         42

5.1 Discussion Of Result                                                                                                                    42

5.2 Conclusion                                                                                                                                    43

5.3 Recommendation                                                                                                                         43

Reference

Appendix

LIST OF TABLES

Table                          Title                                       Pages

4.1            Distribution of student according to age group and sex                                                    37

4.2            Prevalence of UTI in different genders                                                                             38

4.3            Prevalence of UTI in different age groups                                                                        39

4.4           The morphology and biochemical identification of various urinary isolates                       40

4.5           Distribution of isolated uropathogens                                                                                41       

4.6           Distribution of Bacteria Susceptibility to Antibiotics                                                        42

LIST OF FIGURES

Figures                                               Title                                          pages

1               Prevalence of UTI among different groups of people            3

2                Classification of Gram negative organisms implicated in pathogenesis of UTIs              15

3               Trichomonasvaginalis                                                          17

4              Infection routes of UTIs                                                         18

CHAPATER ONE

1.0     INTRODUCTION

Gram-negative bacteria are bacteria that do not retain the crystal violet stain used in the gram-staining method of bacteria differentiation (Baron et al.,1996). They are characterized by their cell envelopes, which are composed of a thin peptidoglycan cell wall sandwiched between an inner cytoplasmic cell membrane and a bacteria outer membrane. The gram-negative bacteria include the model organism Escherichia coli, as well as many pathogenic bacteria, such as Pseudomonas aeruginosa, Neisseria gonorrhoeae, Chlamydia trachomatis, and Yersinia pestis.

They are an important medical challenge, as their outer membrane protects them from many antibiotics (including penicillin); detergents that would normally damage the peptidoglycans of the (inner) cell membrane; and lysozyme, an antimicrobial enzyme produced by animals that forms part of the innate immune system. Additionally, the outer leaflet of this membrane comprises a complex lipopolysaccharide (LPS) whose lipid A component can cause a toxic reaction when these bacteria are lysed by immune cells. This toxic reaction can include fever, an increased respiratory rate, and low blood pressure - a life-threatening condition known as septic shock (Pellitier, 2017).

Gram-negative bacteria display these characteristics: An inner cell membrane is present (cytoplasmic),  thin peptidoglycan layer is present (This is much thicker in gram-positive bacteria),Has outer membrane containing lipopolysaccharides (LPS, which consists of lipid A, core polysaccharide, and O antigen) in its outer leaflet and phospholipids in the inner leaflet, porinsexist in the outer membrane, which act like pores for particular molecules, between the outer membrane and the cytoplasmic membrane there is a space filled with a concentrated gel-like substance called periplasm,the S-layer is directly attached to the outer membrane rather than to the peptidoglycan, if present, flagella have four supporting rings instead of two,teichoic acids or lipoteichoic acids are absent, lipoproteins are attached to the polysaccharide backbone, some contain Braun's lipoprotein, which serves as a link between the outer membrane and the peptidoglycan chain by a covalent bond, most, with very few exceptions, do not form spores (Pellitier, 2017).

Urinary Tract Infection (UTI) remains the most common bacteria infection in human population and is also one of the most frequently occurring nosocomial infections (Gastmeiret al., 1998). Its annual global incidence is of almost 250 million (Ronald, 2001). Worldwide, about 150 million people are diagnosed with UTI each year, costing the global economy in excess of 6 billion US dollars (Akram, 2007). Proliferation of bacteria in the urinary tract is the cause of urinary tract infection. The clinical manifestations of UTI depend on the part of the urinary tract involved, the etiologic organisms, the severity of the infection and the patient’s ability to mount an immune response to it (Foxman, 2003). UTIs are 14 times more common in females than in males. In males the anatomical length of the urethra (20cm) provides a distance barrier that excludes microorganisms from the urinary bladder. Conversely, the short urethra (5cm) in females is more readily transverse by microorganisms. In women, the urethra is much shorter and very close to the anus, which is a constant source of fecal bacteria (Zilevièa, 2005). Bacteria are the primary organisms that cause UTI. Gram positive cause 15-20% and gram negative cause 80-85%. 

UTI is the most frequent nosocomial infection and has been suffering a shift in the etiology and antimicrobial susceptibility, as common as other infection in the last decade. It is important to know the etiology and antibiotic susceptibility of infectious agents to guide the initial empirical treatment (Netoet al., 2003).  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 (Farell , 2003). This study is important for clinicians in order to facilitate the effective treatment and management of patient with symptoms of urinary tract infection.  The inoculums were spread with the wire loop on the media plate. They were incubated aerobically at 37°C for 24 hours (Collee, 1996) After completion of incubation, the inoculated culture plates will be observed for presence of any bacterial growth.

Figure1: Prevalence of UTI among different groups of people

1.2AIMS AND OBJECTIVES

 Ø  To find out the prevalence of gram negative organisms in the urine/urinary tract of Michael Okpara University Of Agriculture, Umudike.

Ø  To investigate their antibiotic sensitivity/susceptibility pattern to enable formulation of drugs for urinary tract infection in our community.

Ø  To determine the age and sex prevalence.

Ø  To determine the prevalence of bacterial strains and their antimicrobial susceptibility in urine.

Ø  To find the pathogenic bacteria commonly responsible with UTI and susceptibility patterns this will help the clinicians to choose the right empirical treatment

Buyers has the right to create dispute within seven (7) days of purchase for 100% refund request when you experience issue with the file received. 

Dispute can only be created when you receive a corrupt file, a wrong file or irregularities in the table of contents and content of the file you received. 

ProjectShelve.com shall either provide the appropriate file within 48hrs or send refund excluding your bank transaction charges. Term and Conditions are applied.

Buyers are expected to confirm that the material you are paying for is available on our website ProjectShelve.com and you have selected the right material, you have also gone through the preliminary pages and it interests you before payment. DO NOT MAKE BANK PAYMENT IF YOUR TOPIC IS NOT ON THE WEBSITE.

In case of payment for a material not available on ProjectShelve.com, the management of ProjectShelve.com has the right to keep your money until you send a topic that is available on our website within 48 hours.

You cannot change topic after receiving material of the topic you ordered and paid for.