COMPARATIVE ANALYSIS OF THE IN VITRO ANTIMICROBIAL ACTIVITIES OF THREE GROUPS OF ANTIBIOTICS AGAINST ESCHERICHIA COLI ISOLATED FROM CLINICAL SPECIMENS

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ABSTRACT  

The in vitro antimicrobial activity of three groups of antibiotics was evaluated against clinical isolates of E.coli. The groups comprised aminoglycosides, fluoroquinolones and the cephalosoprins. The E.coli one hundred (100) in number were isolated from urine, stool, high vaginal swab (HVS), ear swab and wound swab collected from different health care facilities in Umuahia Metropolis. The test was carried out by disc diffusion technique. E.coli from high vagina swab showed 84% susceptibility to streptomycin, 78% to gentamycin, 63% sensitivity to ciprofloxacin and 58% to ofloxacin, it was 68% resistant to ceftriaxone and 58% resistant to cefixime. E.coli from urine showed 84% sensitivity to streptomycin, 58% sensitivity to gentamycin and was 75% ,65%, 58% and 69% resistant to ceftriaxone, cefixime, ciprofloxacin and ofloxacin respectively. E.coli from ear swab was 86%, 71%, 64% sensitive to streptomycin, gentamycin and ciprofloxacin respectively and was 64%, 71% and 50% resistant to ceftriaxone, cefixime and ofloxacin respectively. In stool culture it was 83% sensitive to streptomycin and 67% to gentamycin, it was 100% resistant to ceftriaxone, 67% resistant to cefixime, 83%  resistant to ciprofloxacin and 67% resistant to ofloxacin. In wound swab E.coli was 64% sensitive to streptomycin. 83% to gentamycin and 50% to ofloxacin and it was 100%, 100%, 50% resistant to ceftriaxone, cefixime and ciprofloxacin respectively. Streptomycin was the most effective followed by gentamycin.  






TABLE OF CONTENTS

Title page

Certification                                                                                                                           i

Dedication                                                                                                                              ii

Acknowledgement                                                                                                                  iii

Table of content                                                                                                                      iv

List of tables                                                                                                                           vii

Abstract                                                                                                                                  viii

CHAPTER ONE

1.0 INTRODUCTION                                                                                                            1                                                                                                                                                                                           

1.1 Aim of Study                                                                                                                    2                                                                                                               

1.2 Objective of Study                                                                                                            2

1.3 Limitations                                                                                                                       3

CHAPTER TWO

2.0 Literature review                                                                                                              4

2.1 ANTIBIOTICS                                                                                                                 5

2.1.1 Aminoglycosides                                                                                                           5

2.1.2 Fluoroquinolone                                                                                                            7

2.1.3 Cephalosporin                                                                                                                9

2.2 Antimicrobial Activity                                                                                                     11

2.2.1 Antimicrobial activity in vitro

2.2.2 Antimicrobial activity in vivo                                                                                       12

2.3   Resistance to Antimicrobial Drug

2.4 Factors Affecting Antimicrobial Activity                                                                        12

2.5 Clinical use of Antibiotics                                                                                                12

2.6 Characteristics of Antibiotics                                                                                           14

2.7 Escherchia coli                                                                                                                 15

2.7.1 Morphology                                                                                                                   15

2.7.2 Biochemical characteristics                                                                                          15

 2.7.3 Serotypes                                                                                                                      15

2.7.4 Pathogenicity                                                                                                                 18

CHAPTER THREE

3.0 MATERIALS AND METHODS

3.1 Study Area                                                                                                                        26

3.2 Materials and Reagent                                                                                                      26

3.3 Sample collection                                                                                                             26

3.4 Media Preparation                                                                                                            27

3.5 Inoculation and Isolation                                                                                                  27

3.6 Identification of E. coli                                                                                                                              27

3.6.1 Morphology                                                                                                                   27

3.6.2 Gram Stain Test                                                                                                             28

3.6.3 Biochemical Characterization                                                                                       29

3.6.3.1 Citrate test                                                                                                                   29

3.6.3.2 Methyl Red Test:                                                                                                        30

3.6.3.3 Voges-Proskauer Test                                                                                                 30

3.6.3.4 Motility, Indole, Urease test (MIU)                                                                            31       

3.6.3.5 Triple Sugar Iron Test:                                                                                               32

3.7 Antibiotic Susceptibility Testing                                                                                      32

CHAPTER FOUR

4.0 RESULT                                                                                                                          33

CHAPTER FIVE                                                                                                                 

5.0 DISCUSSION                                                                                                                   37

5.1     CONCLUSION                                                                                                             38

5.2     RECOMMENDATION                                                                                                38

REFERENCES

 

 

 

  

 

 

 

 

LISTS OF TABLES

 

TABLE        Title                                                                  Page

1                    Sources of the isolates                                                                                           34

2                    Morphological and biochemical identification of E. coli                                         34

3                    No of isolates sensitive to the following drugs (%)                                              35

4                    No of isolates resistant to the following drugs (%)                                               36  


 





 

CHAPTER ONE


1.0 INTRODUCTION                                                                                   

 Antimicrobial is an agent that kills microorganisms or inhibits their growth (Online Dictionary). Antimicrobial activity is the ability of these agents to kill or inhibit the growth of the microorganisms.  Antibiotics are the group of medicines that are produced by microorganisms or formulated synthetically; they have dynamic property of inhibiting bacterial growth or completely suppressing the toxic effects of microorganisms. Accessibility of commercially available broad spectrum antibiotics causing multi drug resistance remains a key global health issue (Khan et al., 2011). Antibiotics have been used for decades but especially in Asia region due to frequent administration of antibiotics in humans, common environmentally existing bacteria are quickly becoming resistant to treatment with these drugs (Hawser et al., 2007). Broad spectrum antibiotics have a comprehensive range of coverage that contributes to the effectiveness of these medicines against both gram positive and gram negative bacteria. E. coli is considered lethal and virulent organism as it causes several infections which are intestinal or extra-intestinal and is becoming a big contributor in morbidities and mortalities worldwide (Hammerum and Heuer, 2009).         

E. coli comprises of non-pathogenic commensal isolates that forms part of the normal flora of humans and various animals (Tenaillon et al., 2010). In humans, they are the major aerobic organism residing in the intestine, typically with around 106 to 109 colony forming units per gram of stool (Tenaillon et al., 2010). The organism is also found in soil and water, usually as a result of fecal contamination. Several variants or pathotypes of E. coli have been described that cause infections of the gastrointestinal system (i.e. intestinal pathogenic E. coli) while other pathotypes cause infections outside the gastrointestinal system (i.e. extraintestinal pathogenic E. coli (Croxen and Finlay, 2010). E. coli is the most common cause of urinary tract infections (UTIs) in humans (Foxman, 2010), and is a leading cause of enteric infections and systemic infections (Kaper, 2004). The systemic infections include bacteremia, nosocomial pneumonia, cholecystitis, cholangitis, peritonitis, cellulitis, osteomyelitis, and infectious arthritis. E. coli is also leading cause of neonatal meningitis (Kim, 2012). A wide range of antimicrobial agents effectively inhibit the growth of E. coli. The β-lactams (Cephalosporins), fluoroquinolones and aminoglycosides are often used to treat community and hospital infections due to E. coli (Pitout, 2012). β-lactams (Cephalosporins) disrupt cell wall synthesis by binding to and inhibiting the penicillin-binding proteins essential for transpeptidation and carboxypeptidation reactions in cell wall peptidoglycan synthesis. Fluoroquinolones interfere with DNA supercoiling and promote DNA gyrase-mediated double-stranded DNA. The aminoglycosides bind irreversibly to the 50S subunit of the 70S bacterial ribosomes.


1.1 Aim of Study                                                                                            

The aim of this study is to compare the antimicrobial activity of three groups of antibiotics which are Cephalosporins, Aminoglycosides and the Floroquinolones against E. coli isolated from clinical specimens.


1.2 Objective of Study

The objectives of this study is to:

·       To isolate and identify E. coli from clinical specimens.

·       To determine the antibiotic susceptibility pattern of E. coli against the group of antibiotics mentioned above.

·       To know the group of antibiotics with the best antibacterial activity against E. coli.

 

1.4 Limitations

Availability of typing sera would have helped to determine the various serotypes in different clinical samples.                                                                                                                                                                  

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