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ABSTRACT
Continuous increase in the sale and indiscriminate consumption of packaged drinking water and untreated water types such as borehole and well waters in Nigeria is of public health significance.
Three hundred samples comprising 120 sachets, 60 bottled water brands, 60 borehole and 60 well water samples from five sampling sites in Zaria, North Western Nigeria were analysed microbiologically and physicochemically using standard procedures.
Membrane filter technique was used to detect the presence of bacterial indicators of water quality as well as specific pathogens such as Escherichia coli O157:H7 and Enterococcus spp.
The research was conducted between June 2014 to February 2015 during the wet and dry seasons. Isolated pathogens were characterized conventionally, using MicrogenTM ID Kits and molecularly by Polymerase Chain Reactions (PCR).
The nitrate level in samples of the wells (100%) and borehole (40%) water were above the United States Environmental Protection Agency (USEPA) minimum contamination level of 10mg/l.
Cadmium (Cd) and Lead (Pb) contents of boreholes, wells, and brands of sachet waters were above the minimum permissible limits set by NAFDAC (0.003mg/l and 0.01mg/l respectively).
However, Lead (Pb) was not detected in the bottled water brands sampled. Total coliforms and Escherichia coli were detected in sachet water brands (80%), bottled water brands (20%),
borehole (100%) and well water (100%). Enterococci were recovered from sachet water brands (70%), borehole (100%) and well water (100%).
TABLE OF CONTENTS
Cover Page……………………………………………………………………………………………………….. i
Fly Leaf……………………………………………………………………………………………………………. ii
Title Page………………………………………………………………………………………………………….. iii
Declaration………………………………………………………………………………………………………. iv
Certification………………………………………………………………………………………………………. v
Dedication………………………………………………………………………………………………………… vi
Acknowledgements………………………………………………………………………………………….. vii
Table of Contents……………………………………………………………………………………………. viii
List of Tables…………………………………………………………………………………………………… xii
List of Figures………………………………………………………………………………………………….. xv
List of Plates…………………………………………………………………………………………………… xvi
List of Appendices…………………………………………………………………………………………. xvii
Abstract……………………………………………………………………………………………………….. xviii
CHAPTER ONE………………………………………………………………………………………………. 1
- Statement of Research Problem………………………………………………………………… 2
- Justification for the Study…………………………………………………………………………. 2
- Research hypothesis………………………………………………………………………………….. 3
- Aim………………………………………………………………………………………………………….. 3
- Objectives…………………………………………………………………………………………………. 3
CHAPTER TWO.…………………………………………………………………………………………….. 4
- LITERATURE REVIEW…………………………………………………………………………. 4
- Physicochemical Quality of Drinking Water…………………………………………… 4
- Total dissolved solids (TDS)…………………………………………………………………. 4
- pH……………………………………………………………………………………………………… 4
- Electrical conductivity (EC)………………………………………………………………….. 4
2.1.4 Chlorine and chloride ……………………………………………………….’…………………. 5
- Nitrate and nitrite 5
- Heavy metals………………………………………………………………………………………. 5
- Cadmium……………………………………………………………………………………………… 6
- Lead……………………………………………………………………………………………………. 6
- Iron………………………………………………………………………………………………………. 6
- ……………………………………………………………………………………………. 7
- Microbiological Quality of Drinking Water…………………………………………….. 7
- Coliforms (Total and Faecal Coliforms)……………………………………………………. 7
- Antibiotics Resistance in Bacteria…………………………………………………………… 8
- Water Sources in Zaria……………………………………………………………………………. 9
- Production of Packaged Drinking Water………………………………………………….. 9
- Microorganisms Associated With Packaged and Other Drinking Water Sources 9
- Escherichia coli…………………………………………………………………………………….. 10
- Enterococci…………………………………………………………………………………………… 12
- Enteric viruses……………………………………………………………………………………….. 14
- Current Methods and Emerging Approaches for the Detection and Enumeration of Coliforms in Drinking Water…………… 14
CHAPTER THREE………………………………………………………………………………………… 16
- MATERIALS AND METHODS……………………………………………………………… 16
- The Study Area………………………………………………………………………………………. 16
- Sampling Area………………………………………………………………………………………… 16
- Study Design…………………………………………………………………………………………… 16
- Sample Size…………………………………………………………………………………………….. 16
- Collection of Samples……………………………………………………………………………… 18
- Physicochemical Analysis of Water Samples……………………………………………. 18
- Temperature, pH, electrical conductivity and total dissolved solids……………… 18
- Dissolved oxygen and biological oxygen demand……………………………………… 19
- Nitrate………………………………………………………………………………………………….. 19
- Sulphate……………………………………………………………………………………………….. 20
- Determination of heavy metals in the water samples…………………………………… 20
- Bacteriological Analysis of Water Samples Using Membrane Filter (MF) Technique 21
- Enumeration of Escherichia coli using membrane lactose glucuronide agar (MLGA)… 21
- Enumeration of total coliforms…………………………………………………………………. 22
- Enumeration of faecal coliforms……………………………………………………………… 22
- Enumeration of thermotolerant (faecal) Escherichia coli…………………………….. 23
- Enumeration of enterococci…………………………………………………………………….. 23
- Enumeration of total heterotrophic bacteria or total viable count (TVC)………. 23
- Isolation of Escherichia coli O157:H7 from water Samples………………………… 24
- Biochemical Characterization of Isolates by Conventional Methods…………. 25
- Indole test…………………………………………………………………………………………….. 25
- Methyl red-voges proskaeur (MR-VP) test……………………………………………….. 25
- Citrate test…………………………………………………………………………………………….. 25
- Litmus milk decolorization test to identify Enterococcus spp……………………… 26
- Aesculin hydrolysis test to identify Enterococcus spp………………………………… 26
- Oxidase test…………………………………………………………………………………………… 26
- Biochemical Identification of presumptive Escherichia coll isolates using
Microgen™ GnA-ID Kit………………………………………………………………… 26
- Biochemical Identification of presumptive Enterococcus species isolates using Microgen™ STREP-ID………………… 27
- Serological Identification of Presumptive coli O157……………………………. 28
- Antibiotics Susceptibility Testing…………………………………………………………… 28
- Measurement of inhibition zone diameter…………………………………………………. 29
- Multi-drug resistance index…………………………………………………………………….. 29
- Molecular Characterization of Escherichia coli O157:H7 and Enterococcus
species……………………………………………………………………………………………. 29
- DNA extraction using extraction kit……………………………………………………….. 29
- DNA quantification using NanoDrop……………………………………………………… 29
- DNA amplification and detection……………………………………………………………. 31
Electrophoresis of PCR amplicons 31
- Statistical Analysis…………………………………………………………………………………. 31
CHAPTER FOUR………………………………………………………………………………………….. 34
- RESULTS…………………………………………………………………………………………………. 34
- Physicochemical Properties of the Packaged and other Drinking Water Sources in Zaria, Kaduna State, Nigeria………………… 34
- Heavy Metal Content of the Drinking Water Sources and Packaged Water Sold in Zaria, Kaduna State………………………………44
- Bacteriological Quality of the Packaged and other Drinking Water Types in Zaria Kaduna State, Nigeria…………………………..48
- Effects of seasonal variations on the bacteriological quality of the packaged and other drinking water sources in Zaria………………………………………………………………………. 65
- Biochemical Characterization and Identification of Isolates……………………….. 69
- Antibacterial Susceptibility Profile of Bacteria Isolates……………………………….. 74
- Molecular Characterization of Isolates…………………………………………………….. 82
CHAPTER FIVE……………………………………………………………………………………………. 91
- DISCUSSION…………………………………………………………………………………………… 91
- Physicochemical Parameters of the Packaged and other Drinking Water Sources in Zaria………….91
- Heavy Metal Content of the Packaged and other Drinking Water Sources in Zaria. 94
- Bacteriological Quality of the Packaged and other Drinking Water Sources in Zaria. 96
- Antibacterial Susceptibility Profile of the Bacteria Isolates……………………… 100
- Molecular Characterization of Isolates by the Polymerase Chain Reaction (PCR) Technique………………..102
CHAPTER SIX…………………………………………………………………………………………….. 104
- SUMMARY, CONCLUSION AND RECOMMENDATIONS………………….. 104
- Summary………………………………………………………………………………………………… 104
- Conclusion………………………………………………………………………………………………. 106
6.1 Recommendations………………………………………………………………………………….. 107
REFERENCES……………………………………………………………………………………………. 110
APPENDICES……………………………………………………………………………………………… 117
INTRODUCTION
Water is a simple molecule, consisting of two hydrogen atoms bonded to an oxygen atom with molecular formula H2O (Bassem, 2013).
It is one of the most important chemical substances for the sustenance of life and is vital for all known forms of life on Earth; It constitutes about 75% of the Earth`s surface (UN, 2005; EL-Jakee et al., 2009; Hongyue et al., 2013).
In terms of sheer volume, About 97.5% of all the water on Earth is salt water, only 2.5% of all the water on Earth is fresh water and 98.8% of that fresh water is frozen in Antarctica and Greenland icecaps or lies too deep underground to be accessible; only 1.2% of the Earth’s freshwater is available for withdrawal and human use (Shiklomanov, 2000; UN, 2005; Bassem, 2013).
However, this is continually being polluted by various anthropogenic activities thereby further reducing the available freshwater for human use.
The drinking water of most communities including Zaria in Nigeria is obtained from various sources: boreholes, rivers, streams, and well waters.
Source water contamination poses a risk to public health and increases the cost of drinking water treatment.
The threats posed by deteriorating water quality caused by among other things, the contamination of potable water sources have led the public to seek for alternative potable water sources.
REFERENCES
Chigor, V.N., Umoh, V.J. and Smith, S.I. (2010). Occurrence of Escherichia coli O157:H7 in a river used for fresh produce irrigation in Nigeria. African Journal of Biotechnology, 9(2):178-182.
Clark, C., Price, L., Ahmed, R., Woodward, D., Melito, P. and Rodgers, F. et al. (2003). Characterization of waterborne outbreak associated Campylobacter jejuni, Walkerton, Ontario. Journal of Emerging Infectious Diseases, 9(10):1232- 1241. doi:10.3201/eid0910.020584.
Erah, O.P., Akujieze, C.N. and Oteze, G.E. (2002). The quality of groundwater in Benin city:A baseline study on inorganic chemicals and microbial contaminants of health importance in boreholes and open wells. Tropical Journal of Pharmaceutics Research, 1(2):75-82.
Ezeribe, A., Oshieke, K. and Jauro, A. (2012). physicochemical properties of well water samples from some villages in Nigeria with cases of stained and mottle teeth. Science World Journal, 7(1):1597-1599.
Ferreira, A.C., Morais, P.V., Gomes, C. and DaCosta M.S. (1994). Alterations in total bacteria, iodonitrophenyltetrazolium (INT) positive bacteria and heterotrophic plate counts of bottled mineral water. Canadian Journal of Microbiology, 40:72-77.
Fricker, C., Warden, P. and Eldred, B. (2010). Understanding the cause of false negative β-d-glucuronidase reactions in culture media containing fermentable carbohydrate. Letters in Applied Microbiology, 50(6):547-551. doi:10.1111/j.1472-765x.2010.02834.x.
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