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CHAPTER ONE INTRODUCTION 1.1 Background of Study Water quality analysis of the effluent was run on all thirty of the well washes examined in the 2000–2001-time frames. Water quality of effluent was evaluated in all three types of well washes in Boston and Orlando areas, and the self-serve and in-bays in the Phoenix area. Analysis of variance was examined for facility type, the use of reclaim or the location for the EPA’s thirteen priority metals, and a number of organic compounds and pollutants with characteristics, which would be of concern in well washes.

Water is very swellce but an important commodity in many countries worldwide. The major problems nowadays are severe water contamination and insufficient water resources (Enoh & Christopher, 2015). South Africa is the 30th driest country in the world with an average annual rainfall of 495mm as compared with the world’s average of 1033mm of rainfall. South Africa is said to be a water swellce country, with evaporation losses at three times more than rainfall, therefore great attention must be paid to the efficient management and use of water (Hedden & Cilliers, 2014).

Despite the swellcity of this valuable natural resource, they are polluted everyday due to improper effluent treatment and disposal of polluted wastewater leading to public and environmental concerns. This pollution caused by environmental degradation has now become an alarming problem as a result of rapid and unplanned urbanization, high population growth and unskilled utilisation of water resources (Singh & Singh, 2006, Enoh& 1

Christopher, 2015). Water has been under—valued worldwide and many of the shortages of water emerging around the world are due to failures to value water and the illusion created that water is plentiful (Al-Odwani et al., 2007). It is predicted that in countries in Northern and Southern Africa water availability will decline, impacting on freshwater ecosystems. Twenty five countries are expected to experience water swellcity or water stress over the next 20-30 years (Alemayehu et al., 2012).

In urban areas, the continuous population growth demands that public services such as transportation increase and that includes public and private wells, buses and trucks. As a result of increased number of vehicles wellwash facilities also increases (Rubi-Juarez et al., 2015). Borehole is one of the activities that consume large quantities of water, and results in large amounts of wastewater generated by the well wash facilities containing highly hazardous pollutants, but it has received very scant attention as a potential source of water pollution globally as compared to other high profile environmental threats, such as global warming.

Freshly Water, which has been discharged from domestic discharged domestic sewage is a grey turbid liquid with a dwellings, institutions and commercial establishments characteristic but not with unpleasant smell. Domestic wastewater and well wash together will produce unpleasant smell if kept unaerated due to discharge form manufacturing industries (known a s formation of hydrogen sulphides and amines . Industrial wastewater contains a large number of that such problems are avoided or minimized, attention potentially harmful 2

compounds. As it is discharged should be paid to the management of our aquatic directly into a watercourse, serious damage might result resources and also of the pollutants, which enter them. A to the many forms of life, which inhabit this water. In sensible management strategy will involve analysis of the addition, watercourses utilize by man, either as a source composition of wastewater and the receiving stream of potable water or for washing or bathing would present potential risks of the transmission of a large number of The threat to human lives and aquatic animals due to water related diseases. Over the years, man has polluted incessant discharge of untreated sewage cannot be water in various ways, surface water has been polluted dismissed with a wave hand. Ajayi (1996) gave a situation with industrial wastes, wash off from farm lands and even report as it affects the safety of our environment. For domestic sewage (Ogedengbe and Elutade, 2003). instance, in the report, it was indicated that an estimated. Wastewater is a term used to describe a complex mixture of wastes comprising all water discharged directly from domestic homes, wellwashes, road runoff ,industrial and agricultural wastes. The waste water or effluents gotten from well washes are as a result of removing of dirt and grime from vehicles ,these waste water contain high number of pollutants. Pollutants of concern in well wash effluents include diesel range organics (DROs) like oil and grease, wellbon, asphalt, surfactants, salts, detergents, phosphates, ammonium compounds, heavy metals, acids, organic matter and microorganisms among others (Mazumber and Mukherjee 2011; Lau et al. 2013; Mohamed et al. 2014). Some of these pollutants like oil, grease, detergents, heavy metals and ammonium compounds can

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be directly toxic to aquatic organisms like fish while others like nutrients can cause a shift in ecological balance, should they be improperly managed and discharged (Randles et al. 2007).

Oily wastewater contains toxic components such as phenols, petroleum hydrowellbons and polyaromatic hydrowellbons (PAHs) which are inhibitory to plant and animal growth, mutagenic and wellcinogenic to humans. It has long been established that oil toxicity in aquatic systems is mainly due to the soluble compounds as opposed to dispersed oil droplets. Larval stages of aquatic organisms are particularly vulnerable to soluble toxic components in grease and oil and in lubricants in general, which also tend to accumulate in at the base of aquatic food chains, ultimately reaching human beings as the contaminants move up the food chain (Diphare et al. 2013).These waste water connected to riverine areas or connected to streams or rivers hence contaminates the fresh water ;most human are seen using this water for drinking, washing of clothes and food such as fruits or vegetables. When these vegetables and fruits are consumed by humans ,it can cause a variety of health issues to them such as kidney problems ,high risk of cancers and circulatory problems if they are exposed. This problem continues to grow and expand especially in developing countries ,countries like Nigeria with high population and increased volume of wells resulting in the development and population of several well wash industries thereby creating huge amount of effluent

The results of the water quality testing produced several non-detect values, particularly in regards to the 13 priority pollutants. The average value for sites with levels above minimum detection limits is presented along with the number of sites included in calculating the 4

average. The non-detect value is reported using the procedure quantification limit (PQL) for each of the non-detect results. The number of sites used in determining the mean value is presented for each facility type.

1.2 Aims of study

The aim of this study was to isolate, characterize, identify and determine the susceptibility profile of Salmonella and Shigella spp from well water in Oduduwa University Ipetumodu, Ile-ife, Osun state and to asses the toxicity potential or effects of these effluents on the stream water. 1.3 Objective of Study The objectives of the study were to: (a) Isolate, identify, characterize Salmonella and Shigella spp from well wash effluents. (b) Assess the antibiotics susceptibility pattern of the isolates.

1.4 Limitation of Study

There was unsteady power supply making the process for the study slow and time consuming.

1.5 Statement of Problem

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Water is a crucial resource and with the population increasing and the climate changing, the shortage of freshwater resources worldwide will become more critical (Ghadouani&Coggins, 2011).

CHAPTER TWO

LITERATURE REVIEW

2.1 Wastewater

Wastewater or dirty water from the well wash normally will contain detergent and soaps, grease and gasoline, and other residue from exhaust fumes. In addition, some sludge or muds attach on the vehicles also washes off. Heavy metal usually have the greater density present from various human activities such as refining, mining and electroplating industries (K. W. Lawrence, 2009).The heavy metal normally dissolved in the form of solution and cannot be seen with normal eyes. Normally, well wash water discharge contains heavy metals or other contaminants. As an example, detergents can seriously affect the water quality because it contains some pollutants such as; phosphate, sodium, potassium and other contaminants. Detergents can have toxic effect on a wide variety of marine life. That is why detergent must have their certain measurement to use even the biodegradable ones. Furthermore, from grease oil well it contains higher concentration of lead. Majority of the bacteria found in nature live on dead decaying organic matter as Saprophytes (Peter and George., 1989). The contaminated water therefore has critical impact on all biotic components of the ecosystem and this coil

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affect its use for other purposes. Impurities in water may be floating as suspended matter consisting of materials of greater density than water (Prescott et al., 2008). Though the composition of wastewater

varies depending on the sources, disposal of untreated

wastewater into the environment degrades surface and groundwater water quality which pose threat to human health and wellbeing, as well as to ecosystems. Poor management of wastewater would lead to negative impact of ecosystem services on other beneficiaries like animals.

2.1.1 Microorganisms that can be found in wastewater.

Bacterial indicators, such as Escherichia coli (E. coli) and fecal coliforms, are the required microbial measures of effluents for wastewater-discharge permits in Ohio and elsewhere in the United States. Most bacterial indicators do not cause disease but are used to indicate the possible presence of pathogens transmitted via the fecal-oral route. Sampling for viruses and other pathogens in wastewaters is seldom done and is not required. Bacteria, viruses, and protozoa are three types of microorganisms that can be found in wastewater. Waterborne bacterial pathogens in the United States include species of Salmonella, Shigella, Vibrio, Campylobacter, and pathogen strains of Escherichia coli. Waterborne protozoan pathogens include Cryptosporidium and Giardia. Human enteric viruses, which cause a wide range of diseases and symptoms and are excreted in high numbers in the feces of infected individuals, include enteroviruses, adenoviruses, noroviruses, rotaviruses, and hepatitis A virus. Many wastewater-treatment processes can remove microorganisms, but no process removes all 7

microorganisms from wastewater (Al-Odwani et al., 2007). Enteric viruses include waterborne human pathogenic viruses transmitted through the fecal-oral route. Five groups of enteric viruses were targeted during the study—enteroviruses, adenoviruses, noroviruses, rotaviruses, and hepatitis A virus. Human enteroviruses are among the most commonly detected enteric viruses in contaminated waters (Gregory and others, 2006).

2.1.1.1Bacillus Bacillus spp. are Gram-positive, strictly aerobic or facultatively anaerobic encapsulated bacilli. They have the important feature of producing spores that are exceptionally resistant to unfavourable conditions. Bacillus spp. are classified into the subgroups B. polymyxa, B. subtilis(which includes B. cereus, B. brevis and B. anthracis). Disease may result from the ingestion of the organisms or toxins produced by the organisms. Drinking-water has not been identified as a source of infection of pathogenic Bacillus spp., including Bacillus cereus. Bacillus spp. are often detected in drinking-water supplies, even supplies treated and disinfected by acceptable procedures. This is largely due to the resistance of spores to disinfection processes. Owing to a lack of evidence that waterborne Bacillus spp. Are clinically significant, specific management strategies are not required. 2.1.1.2 Campylobacter Campylobacter spp. are microaerophilic (require decreased oxygen) and capnophilic(require increased wellbon dioxide), Gram-negative, curved spiral rods with a single unsheathed polar flagellum. Campylobacter spp. are one of the most important causes of acute gastroenteritis

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worldwide. Campylobacter jejuni is the most frequently isolated species from patients with acute diarrhea disease. Campylobacter spp. occur in a variety of environments. Wild and domestic animals, especially poultry, wild birds and cattle, are important reservoirs. Pets and other animals may also be reservoirs. Food, including meat and unpasteurized milk, are important sources of Campylobacter infections. Water is also a significant source. The occurrence of the organisms in surface waters has proved to be strongly dependent on rainfall, water temperature and the presence of waterfowl. Contaminated drinking-water supplies have been identified as a source of outbreaks (National Research Council 2004). The number of cases in these outbreaks ranged from a few to several thousand, with sources including unchlorinated or inadequately chlorinated surface water supplies and feacal contamination of water storage reservoirs by wild birds. Contaminated drinking-water supplies have been identified as a significant source of outbreaks of campylobacteriosis. The detection of waterborne outbreaks and cases appears to be increasing. Waterborne transmission has been confirmed by the isolation of the same strains from patients and drinking-water they had consumed. Within a WSP, control measures that can be applied to manage potential risk from Campylobacter spp. include protection of raw water supplies from animal and human waste, adequate treatment and protection of water during distribution. Storages of treated and disinfected water should be protected from bird faeces. Campylobacter spp. Are feacally borne pathogens and are not particularly resistant to

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disinfection. Hence, E.coli (or thermotolerant coliforms) is an appropriate indicator for the presence/absence of Campylobacter spp. in drinking-water supplies. 2.1.1.3 Escherichia coli pathogenic strains Escherichia coli is present in large numbers in the normal intestinal flora of humansand animals, where it generally causes no harm. However, in other parts of the body,E. coli can cause serious disease, such as urinary tract infections, bacteraemia and meningitis. A limited number of enteropathogenic strains can cause acute diarrhoea. Several classes of enteropathogenic E.coli have been identified on the basis of different virulence factors, including

enterohaemorrhagic

E.coli

(EHEC),

enterotoxigenic

E.coli

(ETEC),

enteropathogenic E.coli (EPEC), enteroinvasive E. coli (EIEC),enteroaggregative E.coli (EAEC) and diffusely adherent E.coli (DAEC). EHEC serotypes, such as E. coli O157:H7(Collier et al., 1998) and E. coli O111, cause diarrhoea that ranges from mild and non-bloody to highly bloody, which is indistinguishable from hemorrhagiccolitis. Waterborne transmission of pathogenic E. coli has been well documented for recreational waters and contaminated drinking-water. A well publicized waterborne outbreak of illness caused by E. coli O157:H7 (and Campylobacter jejuni) occurred in the farming community of Walkerton in Ontario, Canada. The outbreak took place in May 2000 and led to 7 deaths and more than 2300 illnesses, O’Connor DR (2002). The drinking-water supply was contaminated by rainwater runoff containing cattle excreta (Hoogenboezem., 2007). There is

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no indication that the response of enteropathogenic strains of E. coli to water treatment and disinfection procedures differs from that of other E. coli. 2.1.1.4 Helicobacter pylori Helicobacter pylori, originally classified as Campylobacter pylori, is a Gram-negative, microaerophilic, spiral-shaped, motile bacterium. There are at least 14 species of Helicobacter, but only H. pylori has been identified as a human pathogen. Although H. pylori is unlikely to grow in the environment, it has been found to survive for 3 weeks in biofilms and up to 20–30 days in surface waters. In a study conducted in the USA, H. pylori was found in the majority of surface water and shallow groundwater samples, Baker KH (1999).The presence of H. pylori was not correlated with the presence of E. coli. Possible

contamination of the environment can be through children with diarrhoea or through vomiting by children as well as adults O’Connor DR (2002). Consumption of contaminated drinking-water has been suggested as a potential source of infection, but further investigation is required to establish any link with waterborne transmission. Humans are the principal source of H. pylori, and the organism is sensitive to oxidizing disinfectants. Hence, control measures that can be applied to protect drinking-water supplies from H. pylori include preventing contamination by human waste and adequate disinfection. 2.1.1.5 Salmonella

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Salmonella spp. belong to the family Enterobacteriaceae. They are motile, Gram negative bacilli that do not ferment lactose, but most produce hydrogen sulfide or gas from wellbohydrate fermentation. It is now considered that this classification is below species level and that there are actually no more than 2–3 species ( Salmonella enterica , Salmonella bongori and Salmonella typhi). All of the enteric pathogens except S. typhi are members of the species S. enterica. Salmonella is spread by the faecal–oral route. Infections with non-typhoidal serovars are primarily associated with person-to-person contact, the consumption of a variety of contaminated foods and exposure to animals. Infection by typhoid species is associated with the consumption of contaminated water or food, with direct person-to person spread being uncommon.

Waterborne typhoid fever outbreaks have devastating public health implications. However, despite their widespread occurrence, non-typhoidal Salmonella spp. rarely cause drinking water-borne outbreaks. Transmission, most commonly involving Salmonella Typhi, has been associated with the consumption of contaminated ground water and surface water supplies. Escherichia coli (or, alternatively, thermotolerant coliforms) is a generally reliable index for Salmonella spp. in drinking-water supplies Ajayi(1996). 2.1.1.6 Vibrio

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Vibrio spp. are small, curved (comma-shaped), Gram-negative bacteria with a single polar flagellum. Species are typed according to their O antigens. There are a number of pathogenic species, including V. cholerae, V. parahaemolyticus and V. vulnificus. Vibrio cholerais the only pathogenic species of significance from freshwater environments. The high numbers required to cause infection make person-to-person contact an unlikely route of transmission. Contamination of water due to poor sanitation is largely responsible for transmission, but this does not fully explain the seasonality of recurrence, and factors other than poor sanitation must play a role. The presence of the pathogenic V. cholera O1 andO139 serotypes in drinking-water supplies is of major public health importance and can have serious health and economic implications in the affected communities(WHO., 1993). 2.1.1.7 Shigella Shigella spp. are Gram-negative, non-spore-forming, non-motile, rod-like members of the family Enterobacteriaceae, which grow in the presence or absence of oxygen. Shigella spp. can cause serious intestinal diseases, including bacillary dysentery. Over 2 million infections occur each year, resulting in about 600 000 deaths, predominantly in developing countries. Most cases of Shigella infection occur in childre...