ISOLATION OF COLIFORMS AND DIFFERENCES AMONG COLIFORM BACTERIA

ABSTRACT

This study investigated the differences in growth and susceptibility pattern of coliform bacteria isolated from fish pond, stream and borehole waters. This was done by determining the presence of coliform bacteria as Escherichia coli, Klebsiella spp and Enterobacter spp, Citrobacter and their antibiotic susceptibility profile using commercially prepared antibiotic discs. Results showed that Escherichia coli 17% was more predominant borehole waters than the stream and fish pond waters. The growth of colifroms at different temperatures depict that E. coli and Citrobacter grew very well at temperature range between 30oC to 45oC whereas Enterobacter sp and Klebsiella sp grew fairly well at thermophilic temperature of 45oC. The susceptibility profile of the isolates to ten antimicrobial agents indicated that majority of the isolates showed little susceptibility and was highly resistant to the following antimicrobial agents (nitrofurantoin, amoxycillin, ampicillin, ciprofloxacin, tetracycline, norbactin/norfloxacin, ofloxacin, cefuroxime and gentamicin). This showed that they exhibit multi-drug resistance pattern which is a common feature of medically important coliform bacteria. None of the water sources met the WHO microbiological standards for drinking water and thus pose a serious health risk to its consumers and users if not properly treated. This study suggests that more check should be put in place in order to maintain the WHO standards for borehole water drilling which recommends that boreholes should be located at least 30m away from latrines and 17m from septic tanks as most of the faecal contamination of pipe waters is caused by the seepage of faecal materials into leaking pipes.

TABLE OF CONTENTS

Title page                                                                                                                    i

Certification                                                                                                                ii

Dedication                                                                                                                  iii

Acknowledgement                                                                                                      iv

Table of contents                                                                                                        v

List of tables                                                                                                               vii

Abstract                                                                                                                      viii

CHAPTER ONE: INTRODUCTION                                                                      1

1.2         Water quality changes                                                                                                        1           

1.3         Water quality challenges                                                                                                    4

1.4       Biological indicators of water quality                                                            6

1.5       Aims and objectives                                                                                       7

CHAPTER TWO: LITERATURE REVIEW                                                          8

2.1       Coliform                                                                                                         8

2.2       Coliform bacteria                                                                                                              8

2.3       Environmental significance                                                                            16

CHAPTER THREE: MATERIALS AND METHODS                                          19

3.1       Collection of water samples                                                                           19

3.2       Growth media used                                                                                         19

3.3       Sterilization                                                                                                    19

3.4       Enumeration of bacteria                                                                                 20

3.5       Identification of bacterial isolates                                                                  20       

3.6       Gram staining                                                                                                 20

3.7       Biochemical cultural characteristics                                                              21

3.7.0    Coagulase test                                                                                                 21

3.7.1    Citrate test                                                                                                       21

3.7.2    Motility, indole, urease test (MIU)                                                                 20

3.7.3    Triple sugar iron test                                                                                       22

3.7.4    Oxidase test                                                                                                    22

 3.8      Antibiotic susceptibility studies                                                                     22

CHAPTER FOUR: RESULTS                                                                                 24

CHAPTER FIVE: DISCUSSION, CONCLUSION AND RECOMMENDATION         

5.1        Discussion                                                                                                 31

5.2       Conclusion                                                                                                      33

5.3       Recommendation                                                                                           33

References                                                                                                      35

LIST OF TABLES

Table                   Title                                                                Page

1:         Total viable count                                                                           25

2:         Biochemical and cultural and morphological characteristics of isolates            26

3:         The total coliform count of bacterial isolates                                                 27

4:         The prevalence of coliform from bacterial isolates                                        28

5:         Growth of coliform bacteria at different temperatures                                  29

6:         Percentage resistance of coliform bacteria isolates from test samples to       30

different antibiotics.

CHAPTER ONE

INTRODUCTION

Water quality is defined in terms of the chemical, physical, and biological characteristics of water, usually in respect to its suitability for an intended purpose (Meybeck et al., 1996). No single measure constitutes good water quality; this is because the quality of water appropriate for recreational purposes differs from that used for industrial processes. Also water suitable for drinking can be used for irrigation, but water used for irrigation may not meet drinking water guidelines (USDA-CSREES, 2001). Water quality is an important determinant of availability because water which is not fit for use is in effect unavailable. The quality and  quantity of available water have implication on the health status of a community. Polluted water is the major reason for the spread of many endemic diseases like skin and eye infections, cholera, tuberculosis, typhoid, diarrhea, viral hepatitis A and even death (Harrison, 1958; Lenat and Crawford, 1994; Biggs, 1995; Gergel  et al.,1999 et al., 2003; Donohue et al., 2006, and Khan et al.,2012 50,000 people die daily due to water borne diseases (Herschy, 1999), also mortality in children less than five years due to water related diseases annually is estimated to be about 4 million in under developed countries. In addition, the World Health Organization estimates that 3.4 million people, mostly children, die every year from water-related disease (WHO, 2004). Ota is the capital of Ado-Odoota Local Government Area in Ogun State, with an estimated 163,783 residents living in or around there. The primary influence on groundwater and surface water quality in Ota is the contamination brought about by human activity. These contaminations include fertilizers and pesticides in agricultural runoff, domestic waste water industrial wastewater, septic tank leachate, and hydrologic modifications. In Nigeria, availability of quality water has become a significant and imperative challenge posing a great concern to families, communities and the Government (Okonko  et al.,2008 Statistics survey from Ota State Hospital from March, 2011 to February, 2012 revealed over 415 cases of water borne diseases and few death cases due to faecal contamination of water. It is therefore important that the quality (physical, chemical and water in Ota be monitored so as to prevent more health hazards in the environment. The ensuring of good quality drinking water is a basic factor in guaranteeing public health, the protection of the environment and sustainable development (Ranjini et al., 2010). Water of good drinking quality is of basic importance to human physiology and man’s continued existence depends very much on its availability (Lemikanra, 1999; FAO, 1997). The provision of portable water to rural and urban population is necessary to prevent health hazards associated with poor drinking water (Nikoladze and Akastal 1989; Lemo, 2002). A significant proportion of the world’s population use potable water for drinking, cooking, personal and home hygiene (WHO, 2004). 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 (Tebutt, 1983). Potable water is defined as water that is free from disease producing microorganisms and chemical substances deleterious to health (Ihekoronye and Ngoddy, 1985).Water is the most common solvent for many substances and it rarely occurs in its pure nature (Caccio, 1973). Water can be obtained from a number of sources, among which are streams, lakes, rivers, ponds, rain, springs andwells (Okonko et al., 2008).Drinking water has always been a major issue in many countries, especially in developing countries like Nigeria (Rajietal., 2010). In Nigeria, majority of the rural populace do not have access to potable water and therefore, depend on well, stream and river water for domestic use (Shittuetal., 2008). Waterof good drinking quality is of basic importance to human physiology and man’s continued existence depends very much on its availability (Lamikanra, 1999; FAO, 1997). The provision of portable water to the rural and urban population is necessary to prevent health hazards (Obi and Okocha, 2007).The failure in government responsibility to provide potable water has made the people of Eyaen community to source for water from other available sources. Supplies are usually derived from springs, rivers, boreholes, streams, and reservoirs. They depend solely on wells and boreholes as their sources, in dry seasons when some of the wells would have dried, many people buy water from water tankers who hawk with their vehicle or fetching from nearby rivers to meet their daily needs. This situation may lead to water borne diseases even as water hawking business is booming in the town. During passage through the ground, water dissolves minerals in rocks, collect suspended particulate matter, particularly those of organic sources as well as pathogenic micro-organisms from faecal matters (Onuh and Isaac, 2009). In some areas, water sources are shared with the animals making the water dirty and contaminated. The quality and quantity of available water have implication on the health status of a community. Over 50,000 people die daily due to water borne diseases (Marque et al., 2003). About 2.3 billion people Worldwide have mortality and morbidity associated with water related ailment. Certain minerals are also toxic such as the heavy metals. Although, some of the heavy metals such as Zinc, manganese, nickel, and copper act as micro-nutrients at lower concentrations, but become toxic at higher concentrations. Health risk due to heavy metal contamination of water through soil has been reported (Eriyamremu et al., 2005; Muchiweti et al., 2006, Singh et al., 2010). Therefore, auditing and monitoring of physic-chemical, minerals, and microbial quality of drinking water as fast becoming an essential aspect of water quality studies. Dirty and polluted water can contain many harmful organisms including pathogenic bacteria, which cause diseases like cholera, bacillary dysentery, typhoid, and diarrhea. Disinfection of water aims to kill these pathogens without leaving any harmful chemical substances in the water. Coliform bacteria, thermotolerant (faecal) coliforms and Escherichia coli have for almost a century been used as indicators of the bacterial safety of drinking water (Leclerc et al., 2001). Water quality guidelines state that drinking water must not contain waterborne pathogens. More specifically, Escherichia coli or thermotolerant coliforms should not be present in any 100 ml sample of drinking water (WHO 2004). The guidelines further state that should this value be exceeded, immediate investigative action must be taken, including repeated testing, thorough inspection of the water source, and the general hygiene of the water distribution system. Unlike other indicators, such as Escherichia coli or total coliforms, low concentrations of heterotrophic plate count (HPC) bacteria will still be present after treatment of drinking water. In general, water purification can achieve heterotrophic bacteria concentrations of 10 colony-forming units (CFU) per milliliter or less in finished water (Fox and Reasoner, 1999).

1.2       WATER QUALITY CHANGES

Natural waters are subject to important changes in their microbial quality. These changes have direct impacts on the decisions made by water authorities striving to maintain safe conditions in catchments or distribution systems. Correct decision making by water authorities relies heavily on having access to rapid and accurate bacteriological data (Daniel et al., 2003). This can be obtained by using  HPC, which is a suitable tool for monitoring changes in bacterial water quality over time for a particular catchment or distribution system (Daniel et al., 2003).Changes in the microbial quality of water may arise from agricultural use, discharges of sewage, wastewater resulting from human activity, and storm or surface water runoff. Previous studies have suggested that sewage effluents contain a wide variety of pathogenic microorganisms whose density and variety are related to the size of the human population, the seasonal incidence of the illness, and dissemination of pathogens within the community (Pipes, 1982). Discharge of domestic sewage into water bodies also depletes dissolved oxygen leading to low dissolved oxygen concentrations and high numbers of enteric bacteria. An improvement of water quality is associated with an increase in the concentration of dissolved oxygen and a decrease in the load of faecal  coliforms. Changes in water conductivity results from changes in the mineral composition of water, which may be caused by seasonal variations in the chemical composition of the various sources of water. It may also indicate sewage, industrial or agricultural pollution or intrusion of saline waters. Determination of various water quality properties on a regular basis may, therefore reveal a need to adjust water treatment according to changes in raw water quality.

1.3       WATER QUALITY CHALLENGES

Livestock practices that can impact on water quality include both intensive and non-intensive operations. Intensive agricultural livestock operations (waste management and disposal) have been identified as point sources of pollution to streams. In water scarce areas, livestock and wildlife density tends to be high in water catchment areas and near water sources due to the presence of pasture and water. These animals generate large quantities of wastes. Water quality changes associated with livestock production include changes in nutrients loads, (nitrogen and phosphorus), microorganisms (e.g. bacteria, faecal coliforms, Cryptosporidium, Giardia) and organic material such as livestock wastes. Localized concentration of animal waste is considered a point source of pollution for surface or ground water. Lack of manure management can adversely affect the water quality of receiving streams, its aquatic life, and reuse of the water downstream for agricultural, recreational and drinking water purposes. High animal densities within the catchment area of a water source may lead to the loss of protective cover of grasses, herbs, and shrubs due to trampling, grazing and browsing action. This exposes the soil to agents of erosion. Increased erosion results in a loss of organic matter, fine soil particles, nutrients, and microbes in the soil (Harper and Marble 1988; Schimel et al., 1985; Belnap, 1995). They may be transported by surface runoff to eventually contaminate drinking water.

1.4       BIOLOGICAL INDICATORS OF WATER QUALITY

The presence of faecal coliforms (over 99 % of which are Escherichia coli) in a water body is     an    indication of possible human/animal waste contamination and the possible presence of pathogenic bacteria. The detection of Escherichia coli provides definite evidence of faecal contamination. However, in practice, the detection of thermo tolerant (faecal) coliform bacteria is an acceptable alternative. According to World Health Organisation (WHO, 1997, 2004) standards, faecal coliforms should be absent (0 colony forming units per 100 ml water) in portable water while total coliforms should be less than 10 colony forming units in any 100 ml water sample. The measurement of faecal coliforms can give an indication of the likely chlorine demand and also indicates where more intensive treatment is needed.

1.5       AIMS AND OBJECTIVES

The aim of this research work is to isolate coliforms from different water sources and to study the differences amongst them.

 OBJECTIVES

1.              To isolate coliforms and determine the differences among coliform bacteria isolated

2.              To determine the antimicrobial sensitivity of the coliform bacteria isolated.

Buyers has the right to create dispute within seven (7) days of purchase for 100% refund request when you experience issue with the file received. 

Dispute can only be created when you receive a corrupt file, a wrong file or irregularities in the table of contents and content of the file you received. 

ProjectShelve.com shall either provide the appropriate file within 48hrs or send refund excluding your bank transaction charges. Term and Conditions are applied.

Buyers are expected to confirm that the material you are paying for is available on our website ProjectShelve.com and you have selected the right material, you have also gone through the preliminary pages and it interests you before payment. DO NOT MAKE BANK PAYMENT IF YOUR TOPIC IS NOT ON THE WEBSITE.

In case of payment for a material not available on ProjectShelve.com, the management of ProjectShelve.com has the right to keep your money until you send a topic that is available on our website within 48 hours.

You cannot change topic after receiving material of the topic you ordered and paid for.