ENUMERATION OF COLIFORMS FROM WELL WATERS LOCATED NEAR SEPTIC TANKS IN ABAKALIKI METROPOLIS, EBONYI STATE



ABSTRACT
Bacteriological examination of well water samples located near septic tanks in Abakaliki metropolis was investigated. 15 well water samples from 15 different wells were used for the research. Measuring tape was used to measure the distance, the colour and physical appearance of the water was observed with the naked eyes, the temperature was measured with mercury thermometer and its pH was measured with pH indicator. Pour plate method were used for the enumeration of viable bacterial cells in the well water while most probable number (MPN) was used for the enumeration of coliforms. Biochemical tests were employed in the identification of. E.coli. The highest distance of 23.17 meters and the shortest distance of 5 metres of the wells from the septic pit were the range used. 

Physical and bacteriological characteristics of the well water samples showed that 9 (60%) had colour ranging from slightly milky to light brown while 6 (40%) is colourless. 10 (66.67%) had presence of particles and cloudy appearance while 5 (33.33%) had no particles in them.The pH of the well waters ranged from 6 to 8 with 10 (66.67) of samples having pH of 7. The temperature range of the well water samples were between 31.80C to 37.00C. Three samples (20%) presented odour. The highest standard plate counts of 2.9x105 cfu/ml were recorded while the lowest count was 3.8x104. The highest most probable number (MPN) per ml was 0.36 and the least was 0.27. Thirteen samples (86.67%) showed the presence of coliforms in them while five samples (33.33%) showed the presence of E coli. It was observed that all the well water sample that give positive E coli came from wells nearest to the septic tank. Well water samples from wells that were 19 meter from the septic tank and beyond did not give positive coliform or E coli. Recovering of E coli from the well water samples may indicate contamination of the well waters from the septic tank. It is therefore recommended that wells water should be sited at least 19 meters from septic tank.       

ENUMERATION OF COLIFORMS FROM WELL WATERS LOCATED NEAR SEPTIC TANKS IN ABAKALIKI METROPOLIS, EBONYI STATE

A PROJECT SUBMITTED TO THE DEPARTMENT OF APPLIED MICROBIOLOGY IN PARTIAL FULFILLMENT OF THE REQUIREMENT FOR THE AWARD OF POSTGRADUATE DIPLOMA (PGD)

                                                  TABLE OF CONTENTS               
Title page                                                                                                       i
Approval Page                                                                                              ii
Certification                                                                                                  iii
Dedication                                                                                                     iv
Acknowledgements                                                                                     v
Abstract                                                                                                         vi
Table of contents                                                                                         vii
List of Table                                                                                                  xi
List of Figures                                                                                               xii
CHAPTER ONE:
1.0.0    INTRODUCTION                                                                          1
1.1.0    Aim of the Research                                                                       4
CHAPTER TWO:
2.0.0               LITERATURE REVIEW                                                  5         
2.1.0               Construction of Well                                                           8
2.2.0               Classification of Well Water                                              10
 2.3.0              Chemical and Physical Properties of Water                    11
2.4.0               Uses of Well Water                                                              14
2.4.1               Agriculture                                                                            16
2.4.2               Drinking                                                                                16
2.4.3               Washing                                                                                 17
2.4.4               Water Industry                                                                      18
2.4.5               Food Processing                                                                   18
2.5.0               Microorganisms Associated with well Water                  19 Contamination                                                               
2.5.1               Coliform Bacteria                                                                20
2.5.2               Giardia Lambia                                                                     21
2.5.3               Crptosporidium                                                                    23       
2.5.4               Hepatitis                                                                                24
2.6.0               Sources of Contamination                                                  26
2.7.0               Effect of Drinking Contaminated Well Water                 26
2.8.0               The septic system                                                                 27
CHAPTER THREE:
3.0                   MATERIAL AND METHODS                                        
3.1.0               Materials                                                                               29
3.1.1               Equipment                                                                             29
3.1.2               Media                                                                                     29
3.1.3               Reagents                                                                                29
3.2.0               Study Area                                                                            29
3.3                   Sample Collection                                                                32
3.4                   Analysis of Physical Characteristics of the Well Water samples 33
3.4.1               Colour and Appearance                                                      33
3.4.2               Determination of Temperature                                          33
3.4.3               Determination of pH                                                            33
3.5.0               Microbiological Analysis                                                   34
3.5.1               Standard Plate Count                                                           34
3.5.2           Serial Dilutions                                                                         34
3.5.3.0            Enumeration of Total Coliforms                                                    35
3.5.3.1            Multiple Tube Method                                                                    35
3.5.3.2            Presumptive Test                                                                              36
3.5.3.3            Confirmed Test                                                                                 36
3.5.3.4            Completed Test                                                                                38
3.5.3.5            Identification of Escherichia coli Colonies on EMB
Agar plate                                                                              39
CHAPTER FOUR:
4.0       RESULTS
4.1       Physical Characteristics of the Well Water Samples                 41
4.2       Standard Plate Count of the Well Water Sample                                    44

4.3       Result of the Presumptive Test and MPN of the Well
Water  Samples                                                                                 46
4.4             Colonial Characteristic of the Organisms on Eosin
Methylene Blue (EMB) agar                                                          48
4.5       Identification of E coli                                                                    50
CHAPTER FIVE
5.0       Discussion                                                                                         52
CHAPTER SIX
Summary, Conclusion and Recommendation
6.1       Summary                                                                                          55
6.2       Conclusion                                                                                        55
6.3       Recommendation                                                                             56
Reference                                                                                         58
Appendix:                                                                                         63

LIST OF TABLES
1:                    Physical characteristics of the Well water
Samples                                                                                 43

2:                    Standard plate count of the Well water samples             45
3:                    Result of the presumptive test and MPN of the
Well water samples                                                              47
4:                    colonial characteristics of the organisms on
                        Eosin methylene blue (EMB) agar                                    49
5:                    Identification of E. coil                                                       51

LIST OF FIGURE

Fig 1: Model of hydrogen bonds between molecules of
                        Water……………………………………………….        3

Fig 2: Map of Ebonyi State showing the study Area…                         31

Fig 3:             Picture showing the growth characteristic and
 colour of Coliforms on  slant Eosin Methylene Blue
(EMB) agar …….. ……………………………………    37

Fig 4:             Picture showing  well distance to septic tank……            70

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