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MICROBIOLOGICAL AND PHYSICOCHEMICAL ANALYSIS OF VARIOUS SACHET AND BOREHOLE WATER CONSUMED IN COMMUNITIES IN OWERRI WEST, IMO STATE
ABSTRACT
This research work was conducted to determine the microbiological and physicochemical state of selected sachet water brands and borehole water consumed in both Nekede and Ihiagwa communities in Owerri West LGA, Imo State, Nigeria, to ascertain the microbiological safety and physicochemical quality of the drinking-water in both communities. Ten commercial borehole water and ten (10) popular sachet water samples were randomly collected from both communities (Nekede and Ihiagwa) in Owerri West and the physicochemical and microbiological properties were analyzed. Twelve bacteria were isolated, of which, three of the isolates were Gram negative bacteria (Chromobacterium sp., Flavobacterium sp. and Pseudomonas sp.), four were non – spore forming Gram positive bacteria (Micrococcus sp., Staphylococcus sp., Corynebacterium sp., and Aerococcus sp.) and five species of spore forming Gram positive bacteria (Bacillus sps.). The physicochemical analysis of the samples (sachet and borehole) indicated that only sachet water samples 5, 9, 10 and borehole water samples 1, 9, 10 were below the WHO permissible limit for pH level (6.5 – 8.5). The temperature range of the samples were 30.00C – 32.10C and 32.10C – 34.50C for sachet and borehole water respectively indicated level above the permissible limit of WHO (270C – 290C). These results obtained for pH and temperature may be attributed to the presence of acidic metabolites and changes in environmental condition. The level of calcium 26.24 – 35.20mg/l and 29.24 – 38.20mg/l, chlorine 68.00 – 82.00mg/l and 74.10 – 94.24mg/l, magnesium 9.45 – 22.10mg/l and 12.50 – 24.10mg/l and hardness 65 – 84mg/l and 70 – 88mg/l for sachet and borehole water samples respectively were within the permissible limit of WHO. The results for Most Probable Number (MPN), Salmonella – Shigella and Vibrio counts indicated that all the samples analyzed recorded zero count for Coliform and Vibrio species suggesting that the samples are not contaminated with fecal material. The result also indicated a higher total culturable heterothrophic bacteria count ranging between (1.2 × 102– 2.9× 102cfu/ml and 1.3 × 102 – 2.4 × 102cfu/ml) for sachet and borehole water samples respectively. The antimicrobial susceptibility of the isolates was determined by disc diffusion method. The antimicrobial profile indicates that most of the Gram positive isolates were intermediate sensitive (≥10mm to ≤15mm) to the tested antibiotics while others were resistant. The Gram negative isolates on the other hand were resistant to most tested antibiotics with zone diameter ≤10mm. Hence, public education should be initiated and intensified on the need to purify water to make it fit for drinking and other domestic purposes.
KEY WORDS: Heterotrophic, Bacteria, Borehole water, Sachet water, Contaminants, Isolates, Water quality
TABLE OF CONTENTS
Certification i
Dedication ii
Acknowledgements iii
Abstract iv
CHAPTER ONE:
1.0 Introduction 1
1.1 Background to the study 1
1.2 Statement of the problem 6
1.3 Objectives of the study 6
1.3.1 General Objective 6
1.3.2 Specific Objectives 7
1.4 Research questions 7
1.5 Significance of the study 7
1.6 Scope of study 8
1.7 Operational Definition of terms 9
CHAPTER TWO: Literature Review
2.1 An Overview 10
2.2 Conceptual Framework 16
2.2.1 Drinking Water Quality 16
2.2.2 Water pollution and Contamination 21
2.2.3 Indicators of Microbial Water Quality 31
2.2.4 Public Drinking water Supply and Sanitation in Nigeria 38
2.2.5 Water Treatment and Household Storage 41
2.2.6 Water Quality Standards 44
2.3 Empirical Studies 49
CHAPTER THREE: Materials and Methods
3.1 Study design 54
3.2 Study Area 54
3.3 Sample and sampling technique 56
3.4 Instruments for Data Collection 56
3.5 Media Used 57
3.6 Sterilization 57
3.7 Diluent Used 57
3.8 Preparation of samples 57
3.9 Analysis of Physical parameters 58
3.10 Culturing of Samples on Various Media 58
3.11 Most Probable Number Multiple Tube Technique 59
3.12 Isolation and Identification of Isolates 60
3.13 Gram Staining 60
3.14 Spore staining 61
3.15 Motility test 61
3.16 Preparation of Standard Inoculum 62
3.17 Biochemical tests 62
3.18 Stock Cultures 65
3.19 Antimicrobial Sensitivity test 66
3.20 Preparation of Inoculum 66
3.21 Antibiotic Sensitivity Disc 66
3.22 Determination of Isolate Susceptibility 67
3.23 Method of Data Collection 67
CHAPTER FOUR:
4.0 Results 69
4.1 Physicochemical Parameters of Sachet Water Samples from Nekede 70
4.2 Physicochemical Parameters of Sachet Water Samples from Ihiagwa 72
4.3 Physicochemical Parameters of Borehole Water Samples from Nekede 76
4.4 Physicochemical Parameters of Borehole Water Samples from Ihiagwa 80
4.5 Population Density of Bacteria from Sachet and Borehole Water Samples 84
4.6 Biochemical Characterization of Bacterial Isolates 89
4.7 Antibiotic Sensitivity Results of the Test Organisms 92 CHAPTER FIVE: Discussion, Conclusion and Recommendations
| 5.1 Discussion | 98 | |||||
| 5.2 Conclusion | 103 | |||||
| 5.3 Recommendations | 103 | |||||
| 5.4 Suggestion for Future Research | 104 | |||||
| REFERENCES | 105 | |||||
| Appendix 1: Field Photographs | 116 | |||||
| Appendix 2: Gram Positive Antibiotics Disc | 122 | |||||
| Appendix 3: Gram Negative Antibiotics Disc | 123 | |||||
| Appendix 4: Bacteria Count of Samples Obtained From Sachet Water | 124 | |||||
| Appendix 5: Bacteria Count Of Samples Obtained From Borehole Water | 125 | |||||
| Appendix 6: Susceptibility Results of Isolates to Various Tested Antibiotics | 126 | |||||
Appendix 7: Susceptibility Results of Gram Positive Isolates to Various Tested
Antibiotics 127
Appendix 8: Susceptibility Results of Gram Negative Isolates to Various Tested
Antibiotics 128
Appendix 9: Sensitivity Zone Diameter and Classification 129
CHAPTER ONE
1.0 Introduction 1.1 Background to the Study
Water is a transparent fluid that forms the world’s streams, lakes, oceans and rain, and it is the major constituent of the fluids of organisms. It covers about 70% of earth’s surface and makes up about 70% of the human body mass and is essential for life (Shakhashiri, 2011). Water is an essential requirement of all life forms. Satisfactory supply of clean, safe, and hygienic drinking water is imperative for health. It is one of the most demanded of all urban and rural amenities and it is indispensable for man’s activities.
Access to safe drinking water is a vital agent for human living (Khatoon and Pirzada, 2010). Unavailability of good quality drinking water is wide spread and this has serious health implication(Onweluzo and Akuagbazie, 2010). Water is one of the most important natural resources known on earth (Ojo, Bakare and Babatunde, 2005).
Water of good quality is important to human physiology and man’s continued existence depends very much on its availability. Before water can be described as potable, it has to comply with certain physical, chemical and microbiological standards, which are designed to ensure that the water is potable and safe for drinking. Potable water is defined as water that is free from contaminants, such as disease causing microorganisms and harmful chemical substances. The importance of potable water supply in the socio-economic life of the public cannot be over emphasized. Often, source and potability of water supply reflects on the health conditions of communities. Water meant for consumption should be free from pollution, acceptable and safe.
Indeed the quality of the water sources should not exceed the maximum limits specified in the water quality guidelines. Water is indispensable for life(Obi et al., 2004).
However, it is estimated that about 1.2 billion individuals worldwide do not have access to potable water (Third World Water Forum on Water, 2003). In many developing countries, availability of water has become a critical and urgent problem and it is a matter of great concern to families and communities that depend on non-public water supply system (Okonko, Ogunjobi, Adejoye, Ogunnusi and Olasogba, 2008).
Adequate supply of fresh and clean drinking water is a basic need for all human beings. Water consumers are frequently unaware of the potential health risks associated with exposure to waterborne contaminants which have often led to diseases like diarrhea, cholera, dysentery, typhoid fever, legionnaire’s disease and parasitic diseases. The inadequacy of pipe borne water supply in Nigeria is a growing problem; as a resort to buying water from vendors, and sachet or bottled water became a major source of drinking water. Although, potable and affordable, the problems of its purity and other health concerns have begun to manifest. Sachet water has been reported to contain bacteria such as Bacillus sp., Pseudomonas sp., Klebsiella sp., Streptococcus sp., and oocysts of Cryptosporidia sp. Apart from environmental contaminants, improper storage and handling by vendors also poses a serious threat to the health of the ignorant consumers (Omalu et al., 2010).
Sachet drinking water was introduced into the Nigerian markets as a less expensive means of accessing drinking water than bottled water (Ogundipe, 2008). There has been countless number of disease outbreak and poisoning around the world resulting from the consumption of untreated or poorly treated drinking water (Fong et al., 2007).
Several studies on the quality of sachet water have reported violation of international quality standards. In Nigeria, small nylon sachets which are electrically heated and sealed at both ends are used to package water; 0.5 liters of water and these were introduced into the market. There are many different brands of sachet drinking water that are beautifully packaged, properly labeled and advertised (Ekwunife, et al., 2010). Although these products are popularly termed
“Pure Water”, they are usually not free of microorganisms (Omemu, Edema and Bankole, 2005;
Taura et al., 2005; Ezeugwunne et al., 2009; Oladipo, Onyenike and Adebiyi,2009). Occasionally, contamination of sachet water may occur either during the processing, transportation or improper handling by hawkers.
Moreover, a greater proportion of the water that is used for the production of sachet water is obtained from boreholes that are exposed to microbial contamination through rainfall runoffs and the fact that they are usually constructed very close to pit toilets. Water related diseases continue to be one of the major health problems globally. The high prevalence of diarrhea among children and infants can be traced to the use of unsafe water and unhygienic practices (Onifade and Ilori, 2008).
In developing countries, 80% of all diseases and over 30% of deaths are related to drinking water (Onweluzo and Akuagbazie, 2010).
Sachet or packaged drinking water is any water that is in a sealed plastic or polythene bag and is distributed or offered for sale and is intended for human consumption (Tagoe et al., 2011). The sale and consumption of packaged drinking water continues to grow rapidly in most of the developing countries of the world (Mgbakor et al., 2011).In the bid to safeguard public health, it is essential that the available packaged sachet water is of highest quality (Anunobi et al., 2006).
The continuous proliferation of this sachet drinking water and their indiscriminate consumption are of concern to public health. An understanding of their microbiological quality and safety are therefore pertinent (Muazu et al., 2012). Microbiological quality of drinking water is defined by the range of microbial variables or microbial parameters present in water, which limit the use of water, while the physicochemical quality of drinking water involves the analysis of the physical and chemical parameters of the drinking water.
Borehole water is extracted from a borehole which is a long, narrow well drilled to access underground water. Before a borehole can be built, technicians and community leaders must work together to find the optimal location and drill a test borehole to assure the quality of water. Pollutants in groundwater can be from various sources mainly municipal (i.e. leakages from liquid waste and solid waste from land fill) industrial (i.e. liquid waste tanks and pipeline leakages, oil field and brines) and agricultural sources (i.e. irrigation return flow which are sometimes saline). These problems of acute water supply have resulted in the rapid increase or hand dug wells and boreholes with some located within the proximity of suck away and pit
latrines.
Groundwater has been naturally very clean because of its filtering effects; however, it can be polluted with nutrients and toxic chemicals when surface water carrying these substances drains into groundwater environment. A groundwater aquifer may be as little as 30m from the surface or as much as 300m. Although, it costs more to pump water from deeper aquifer, the water quality in deeper ones are better than the shallow ones since contaminants which the water may be carrying are removed as the water moves through the rocks (United States Environmental
Protection Agency [USEPA] , 2001).
Groundwater is any water that lies in aquifers beneath the land surface, while some of the water which falls as precipitation is channeled into streams or lakes, and some are used by plants or evaporates back into the atmosphere, most of it seeps underground (Oskin, 2015).
The quality of groundwater resource depends on the management of human waste as well as the natural physical characteristics of the catchment areas (Saba and Baba, 2004). Bacteria and their associated colonies account for 80% of the fouling problems found in well systems.
Generally, borehole water is considered to have better microbial quality because its water is from deep aquifer. It contains no or low levels of harmful pathogens but it can be polluted with naturally occurring chemicals. Contamination can be caused by poor sanitary protection at the top of the borehole (World Health Organization [WHO] , 2011).
Several studies has documented the detection of coliform and heterotrophic bacteria in sachet drinking water which far exceeded the national and international standards set for potable water for human consumption (Khaniki et al., 2010). Coliform are commonly used bacterial indicator of sanitary quality of foods and water. They are defined as rod-shaped Gram-negative non-spore forming and motile or non-motile bacteria which can ferment lactose with the production of acid and gas when incubated at 35oC-37oC (American Public Health Association [APHA] , 1995).
Heterotrophic bacteria are non-coliform species of bacteria that utilizes organic substance for its development. The presence of heterotrophic bacteria in drinking water is not an indication that the water presents health risk but poses significant health risk in immunocompromised individuals. Similarly, total coliform bacteria may cause health risk for infants, young children, and elderly people and especially to the immunocompromised individual (Timilshina et al., 2012).
1.2 Statement of the Problem
Water is everywhere but potable water that is safe and suitable for drinking and other human activities can be said to be available, but not evenly distributed. Water-borne diseases due to unsafe water consumption such as diarrhoea, cholera, dysentery, typhoid, etc, have affected and are still affecting public health, especially that of children in most developing countries of the world. Approximately, 88% of deaths due to diarrhoeal illnesses worldwide are attributable to unsafe water consumption, inadequate sanitation and poor hygiene (World Health Organization [WHO] , 2015). It is the second leading cause of death among children under five, as it kills more children than AIDS, malaria and measles combined (World Health Organization [WHO] , 2015).
Therefore, this research work seeks to examine the microbiological safety and the physicochemical quality of drinking water (i.e. the Sachet and Borehole water) consumed in
Ihiagwa and Nekede communities; to determine the safety value of the water they consume. (
1.3 Objectives of the Study
1.3.1 General Objective
The general objective of this study is to determine the microbiological and physicochemical quality of sachet and borehole water samples obtained from selected sachet water brands and borehole locations respectively, in Ihiagwa and Nekede communities in Owerri West, Imo state, Nigeria.
1.3.2 Specific Objectives
In other to achieve the study general objective, the following are the specific objectives of this study:
- To determine microbial load present in the water samples.
- To determine the level of some physicochemical parameters in the water samples.
- To determine the biochemical characteristics of the microbial isolates from the sachet and borehole water samples.
- To determine the antimicrobial sensitivity or characteristics of the isolates.
1.4 Research Questions
- Are there any microbial loads present in both water samples?
- What are the levels of some of the physicochemical parameters in the water samples?
- What are the biochemical characteristics of the isolated microorganisms from the water samples?
- What are the isolates’ antimicrobial sensitivity patterns or characteristics?
1.5 Significance of Study
The significance of this study is to ascertain the physicochemical and microbiological quality of selected sachet water brands and borehole water sources in Nekede and Ihiagwa communities in Owerri west, Imo State Nigeria. The major activity is the necessity to provide data on the level of physicochemical and microbiological properties in the sachet and borehole water samples from both communities. It will also give a summary on the cause and sources of microbiological contamination, if available, with the aim of raising standards in water consumption and also to reassure the public on the safety of the water they consume.
1.6 Scope of Study
The two communities under study have numerous brands of sachet water and borehole water supplies, therefore, the scope of this research covers microbial and some important physicochemical water quality analysis on sachet and borehole water samples randomly selected and collected from sachet water vendors, retail shops where the sachet (packaged) water are sold, from the various sachet water companies and borehole sites, located in the communities, respectively.
This choice of selection is due to financial constraint which could be experienced due to cost of carrying out analysis on these water samples. This study will also be delimited to Ihiagwa and Nekede communities in Owerri West, in Imo State, Nigeria, where the water samples will be collected and this will represent the study area. The research work is also limited to laboratory analysis carried out on water samples to obtain the required results.
1.7 Operational Definition of Terms
- Borehole Water: is extracted from a borehole which is a long, narrow well drilled to access underground water.
- Coliform: are commonly used bacterial indicator of sanitary quality of foods and water. They are rod-shaped, gram-negative, non-spore forming and motile or non-motile group of bacteria which can ferment lactose with the production of acid and gas when incubated at 35oC-37o
- Groundwater: is any water that lies in aquifers beneath the land surface, while some of the water falls as precipitation is channeled into streams or lakes, and some is used by plants or evaporates back into the atmosphere, most of it seeps underground.
- Heterotrophic Bacteria: are non-coliform species of bacteria that utilizes organic substance for its development.
- Microbiological Quality of Drinking Water: is defined by the range of microbial variables or microbial parameters present in water, which limit the use of water.
- Potable Water: is defined as water that is free from contaminants, such as disease causing microorganisms and harmful chemical substances.
- Sachet or Packaged Drinking Water: is any water that is in a sealed plastic or polythene bag and is distributed or offered for sale and is intended for human consumption.
- Water: is a transparent fluid that forms the world’s streams, lakes, oceans and rain, and is the major constituents of the fluids of organisms. It covers about 70% of earth’s surface and makes up about 70% of the human body mass and is essential for life.
MICROBIOLOGICAL AND PHYSICOCHEMICAL ANALYSIS OF VARIOUS SACHET AND BOREHOLE WATER CONSUMED IN COMMUNITIES IN OWERRI WEST, IMO STATE
