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ASSESSMENT OF Escherichia coli FROM URINE OF UNIVERSITY STUDENTS FOR BIOFILM PRODUCTION

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Table of contents

CHAPTER ONE                                                                                                                                1

1.1      Introduction                                                                                     1

1.2      Aims and Objectives                                                                        6

1.3      Objective of the study                                                                      6

1.4      Literature Review                                                                             7

CHAPTER TWO: Materials and Methods                                                   10

2.1      Samples Collection                                                                          10

2.2      Sterilization of Materials                                                                  10

2.3      Preparation of Culture Media                                                            10

2.4      Isolation of Escherichia Coli                                                            11

2.5      Characterization and Identification of Bacteria                                  11

2.5.1   Cultural and Morphological Characteristics                                       12

2.5.2   Gram Stain Reaction                                                                        12

2.6      Identification Methods                                                                     13

2.6.1   Urease Test                                                                                      13

2.6.2   Indole Test                                                                                      13

2.6.3  Oxidase Test                                                                                    14

2.6.4   Catalase Test                                                                                   14

2.6.5   Citrate Utilization Test                                                                     14

2.6.6   Motility Test                                                                                    15

2.6.7   Voges-Proskauer (VP) Test                                                              15

2.7      Biofilm Formation Assay of Escherichia coli                                    16

2.7.1   Test Tube Method                                                                            16

2.7.2   Tissue culture Plate Method                                                              16       

CHAPTER THREE                                                                                    18

3.1      Results                                                                                            18

CHAPTER FOUR: Discussion and Conclusion                                           28

4.1      Discussion                                                                                       28

4.2      Conclusion and Recommendations                                                   30

References                                                                                                 32

 

LIST OF TABLES

List    Title                                                                                               Page

3.1      Isolation rate of Escherichia coli and Other Microorganisms       

          Isolated from Urine Samples                                                            20

3.2      Biochemical Characterization of Bacterial Isolated from Urine            

          Samples                                                                                          21

3.3      Results of Test Tube Method used for Assessment of E. coli for          

          Biofilm Production                                                                          22

3.4      Results of Biofilm Production of different E. coli by Microtiter Plate 

          Method                                                                                           23

3.5      Correlation between Biofilm Produced by E. coli with Microtiter 

          Plate Method and Tube method                                                        24

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 



 

LIST OF FIGURES

List    Title                                                                                                Page

3.1      Typical isolate of E. coli showing metallic sheen colonies on Eosin 

          Methylene Blue agar                                                                        25

3.2      Screening of E. coli for Biofilm Production by Test Tube Method         26

3.3       Screening of E. coli for Biofilm Production by Microtiter Plate Method            27

Abstract

        Escherichia coli is one of the most frequent pathogenic bacteria isolated from urine samples and they are able to attach to the bladder wall and form a biofilm that contribute to virulence of bacteria which resist the body’s immune response. This study was conducted to assess Escherichia coli from urine of university students for biofilm production. 60 urine samples were collected from students of Olabisi Onabanjo University into a sterile universal bottle and transported to the Department of Microbiology laboratory for immediate culture on Eosin Methylene Blue agar. The bacteria isolates were examined and characterized biochemically using standard microbiological method. E. coli was found to be the most frequent isolated pathogen (n=12, 20%), followed by Klebsiella spp. (n=10, 16.6%), E. coli / P. aeruginosa (n=6, 10%), Pseudomonas aeruginosa (n=5, 8.3%) and Proteus mirabilis (n=3, 5%). Out of the Eighteen (18) E. coli subjected to biofilm production, 16 (88.8%) isolates produced biofilm by TCP method and 13(72.2%) produced biofilm by Tube Method. Presence of this pathogen in urine tract may lead to multiplication and establishment of urinary tract infection which females are more prone to than males. The ability of E. coli to form a biofilm which resists the body’s immune response may pose a public health problem for the host patients as the microorganisms in biofilm contribute to virulence of bacteria that are difficult to treat with antimicrobial agents. Good knowledge and proper education should be made known to the students and general populace on urinary tract infections and the causes.

 

Keyword: Escherichia coli, Biofilm formation, Urine

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