ISOLATION, CHARACTERIZATION AND SUSCEPTIBILITY PATTERN OF BACTERIA ISOLATED FROM URINE IN FEMALES OF REPRODUCTIVE AGE IN UMUAHIA METROPOLIS

ABSTRACT

This study was carried out to isolate, characterize and to ascertain the susceptibility pattern of bacteria from urine in female reproductive age in Umuahia metropolis. The streaking method of inoculation was used to isolate the organisms and the disc diffusion method was used for the sensitivity testing. Four bacteria genera were isolated namely: Escherichia coli (40%), Staphylococcus aureus (30%),  Klebsiella pneumoniae (10%), Pseudomonas aeruginosa (10%). E.coli was susceptible to Ampicillin and Gentamicin (20mm respectively), Streptomycin (14mm), Amoxillin (15mm) and Ciprofloxacin (17mm) but resistance to the rest of the drug. Klebsiella pneumoniae was sensitive to only Gentamicin and Ciprofloxacin (15mm and 16mm respectively). Pseudomonas aeruginosa  was sensitive to Ampicillin and Gentamicin (15mm and 14 mm respectively). All the drugs tested exhibited and inhibitory effect against S. aureus. Empirical antimicrobial therapy, indepth knowledge of the etiology, predisposing factors, cultural positivity is essential to avoid negative drug and to establish the optimum prophylactic therapy.

TABLE OF CONTENT

Title                                                                                                                             i

Certification                                                                                                                ii

Dedication                                                                                                                  iii

Acknowledgements                                                                                                    iv

Table of Contents                                                                                                       v

List of Tables                                                                                                              vii

Abstract                                                                                                                      viii

CHAPTERONE: INTRODUCTION                                                                     1                                        

1.2 Aims and objectives of the Study                                                                         4

CHAPTER TWO: LITERATURE REVIEW                                                       

1.0      Introduction                                                                                                       5

2.1 Urinary Tract Infection                                                                                         5

2.2 Classification Of Urinary Tract Infection                                                             7

2.3 Symptoms Of Urinary Tract Infection                                                                 8

1.3.1        Predisposing Factor                                                                                        9

1.3.2        Causative Organisms                                                                                      9

CHAPTER THREE: MATERIALS AND METHODS

3.1 Sample Collection                                                                                                 11

3.2 Transportation Of Urine Sample                                                                           11

3.3 Preparation Of Culture Media                                                                              11

3.4 Isolation Of Bacteria                                                                                            11

3.5 Characterization and identification of pathogens                                                 12

3.5.1 Cultural Observation                                                                                                      12

3.5.2 Gram Staining                                                                                                                12

3.6 Antimicrobial Sensitivity Testing (Kirby-Bauer Method)                                               13

CHAPTER FOUR: RESULT                                                                                             14

Table 1.0 Colony Morphology And Characteristics Of The Isolate                                      15

Table 2.0 Frequency of occurrence of isolate                                                                        16

Table 3.0 Antimicrobial susceptibility pattern in gram negative isolates                               17       

Table 4.0 Antimicrobial susceptibility pattern in gram positive isolates                                18

CHAPTER FIVE: DISCUSSION, CONCLUSION AND RECOMMENDATION

5.1       Discussion                                                                                                                   19

5.2       Conclusion                                                                                                                  20

5.3 Recommendation                                                                                                             20

References                                                                                                                              21

Appendix                                                                                                                                25

LISTS OF TABLES

Colony morphology and characteristics of the isolate                                    Table 1.0

Frequency of occurrence of isolate                                                                 Table 2.0

Antimicrobial susceptibility pattern in gram negative isolates                        Table 3.0

Antimicrobial susceptibility pattern in gram positive isolates             Table 4.0

CHAPTER ONE

1.0     INTRODUCTION

Urinary Tract Infection (UTI) is caused by pathogenic invasion of the urinary tract which leads to an inflammatory response of the urothelium. Proliferation of bacteria in the urinary tract is the cause of urinary tract infection (Haider et al., 2010).

Urinary Tract Infections  (UTIs) are serious health affecting problems worldwide. Escherichia coli, E. faecalis, K. pneumoniae, S. marcescens, P. aeruginosa, S. saprophyticus, S. aureus and Proteus mirabilis are most common bacteria causing UTIs in human beings.

 The E. coli accounts for approximately 85% of community acquired UTIs and 50% of hospital acquired UTIs (Ramanath et al., 2011). Different factors like age, gender, immune-suppression and urological instruments may affect prevalence of UTIs (Igbal et al., 2010). Catheter-associated UTIs are one of the most dangerous health risks contributing 34% of all health care associated infections (Fink et al., 2012).

Urinary Tract Infections  (UTIs) are the most frequent infections which cause pain, fever, discomfort and extra intestinal infections in all age groups of people (Kunin, 1994).

Every year approximately 150 million people are suffering with UTI all over the world. It has been expected that about six million patient visit outpatient departments and about 300,000 are treated in the wards every year for UTI worldwide. About 10% of human population gets UTI at some stage during their lives (Gupta et al., 2001).

Most common causative bacteria is E. coli and more prone to play a role incausing 80-90% of lower UTI, whereas in about 95% of patients suffering from acute pyelonephritis, normally the infecting organisms are Gram negative isolates (Zahera et al., 2011).

The clinical manifestations of UTI depend on the portion of the urinary tract involved, the etiologic organism, the severity of the infection and the patient’s ability to mount an immune system to it. Signs and symptoms may include fever, chills, dysuria, urinary urgency, frequency and cloudy or malodorous urine. Infections are almost always ascending in origin and caused by bacteria in the periurethral flora and the distal urethra. These bacteria inhabit the distal gastrointestinal tract and colonize the perineal area. E. coli usually causes a child’s first infection but other Gram-negative bacilli and Enterococci may also cause infection (Oladeinde et al., 2011). Staphylococcal infections especially those due to Staphylococcus saprophyticus are common causes of urinary tract infection among female adolescents (  Shalini et al., 2011).

Under normal circumstances the kidney, uterus and the urinary bladder of mammals are sterile. Urine within the urine bladder is also sterile. However in both female and male, a few bacteria are usually present in the distal portion of the urethra. The factors for sterility are urine kills some bacteria due to its low pH and the presence of urea and other metabolic end products and much enzymes. The kidney medulla is so hypertonic that few organisms can survive. The lower urinary tract is flushed with urine and some mucus 4-5 times each day, eliminating potential pathogens. In males, the anatomical length of the urethra (20 cm) provides a distance barrier that excludes microorganisms from the urinary bladder. Conversely, the short urethra (5 cm) in females is more readily transverse by microorganisms; this explains why general urinary tract infections are 14 times more common in females than in males. Under the influence of oestrogen, the vaginal epithelium produces increased amount of glycogen that acid-tolerant Lactobacillus acidophilus bacteria called Doderlein’s bacilli degrade to form lactic acid. Normal vaginal secretions contain up to 10⁸ Doderlein’s bacilli per ml thus an acidic environment (pH 3.5) unfavourable to most organisms established. Cervical mucus also has some antibacterial activity. Most UTIs are thought to be caused by organisms originating from the patient’s own bowel.

Urinary tract infection is one of the most frequent occurring nosocomial infections. Normally, UTIs are caused by a variety of Gram-negative and Gram-positive bacteria. The Gram positive bacteria includes Staphylococcus sp, Streptococcus sp and Enterococcus sp. Gram negative includes a large number of aerobic bacilli such as Escherichia sp, serratia sp, Salmonella sp, Citrobacter sp, Proteus sp and Pseudomonas sp. Among this 80-90% of UTI is caused by E. coli and in ambulatory patients and of nocosomial infection, Staphylococcus aureus, Klebseila pneumonia, Enterococcus faecalis, Proteus mirabilis are the most frequently isolated (Boye et al, 2012).

            The pathogenesis of UTIS involves complex interaction between an organism, the environment and the potential host. The symptoms of a person with urinary tract infection depend on the age and the location. (Humayun et al., 2012).

Chronic and acute infection of urinary tract leads to high blood pressure, kidney damage and results in death. Chronic manifestations of the UTIs are acute and chronic pyelonephritis ( a disease process resulting from the effect of infection of parenchyma and pelvis of the kidney), cystitis, renal carbuncle, urethritis and prostatitis. (Rahem et al., 2013).

Treatment of UTIs depends upon sensitivity of bacteria towards a variety of antibiotics such as Trimethoprim-Sulfamethoxazole (TMP-SMX). However, prolonged use of antibiotic cause side effects in the patients and the pathogens develop resistance through plasmid and or mutational change due to course (Manjunath et al, 2011) the sequence of steps in performing a complex study of micro organisms in urine sample includes Aseptic collection of specimens, quantitative analysis, isolation of pathogens, identification of pathogens and Antimicrobial sensitivity testing (Barate et al.,2012).

Treatment of UTIs depends on the bacteria responsible for infection because all the bacteria possess their own susceptibility towards a variety of antibiotics (Ahmad, 2012).

1.2   AIMS AND OBJECTIVES OF THE STUDY

The main aim of this work is to isolate, characterize and ascertain the susceptibility pattern of bacteria isolated from urine samples of female of reproductive age in Umuahia metropolis. The following specific objectives will be determined.

1.      To isolate and characterize the bacteria associated with urine of female of reproductive age.

2.      To determine the susceptibility pattern of each of the isolates against some commercially available antibiotics.

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