Background of Study

Water is a very essential need for man’s survival. Over the years, man has discovered various sources of water. Depending on the characteristics of his geographical area, he has also been tasked to invent suitable and convenient means to assess it.

Water is the universal solvent capable of dissolving nearly all solutes, which is important to living and non-living things [1]. Water is a resource that has many uses, including recreation, transportation, and hydroelectric power, as well as domestic, industrial, and commercial uses [2]. The water of rivers plays an important role in the development of the countries. Water is extraordinarily abundant on the planet as a whole, but fresh potable water is not always available at the right time or in the right place for human or ecosystem use [3].

According to the Encyclopaedia Britannica, surface water and groundwater are both important sources for community water supply needs. Groundwater is a common source for single homes and small towns, and rivers and lakes are the usual sources for large cities. Although approximately ninety-eight percent of liquid fresh water exists as groundwater, much of it occurs very deep in the Earth. This makes pumping very expensive, preventing the full development and use of all groundwater resources.

Water is in constant circulation, powered by the energy from sunlight and gravity in a natural process called the hydrologic cycle. Water evaporates from the ocean and land surfaces, is held temporarily as vapor in the atmosphere, and falls back to the Earth’s surface as precipitation. Surface water is the residue of precipitation and melted snow, called runoff. Where the average rate of precipitation exceeds the rate at which runoff seeps into the soil, evaporates, or is absorbed by vegetation, bodies of surface water such as streams, rivers, and lakes are formed. Water that infiltrates the Earth’s surface becomes groundwater, slowly seeping downward into extensive layers of porous soil and rock called aquifers. Under the pull of gravity, groundwater flows slowly and steadily through the aquifer. In low areas it emerges in springs and streams. Both surface water and groundwater eventually return to the ocean, where evaporation replenishes the supply of atmospheric water vapor. Winds carry the moist air over land, precipitation occurs, and the hydrologic cycle continues.

Subsequently, the quality of water used by man determines his survival too. The quality of water assessable to man depends on nature and human activities. Water quality is defined as the set of physical, chemical, and biological characteristics that must be satisfied in order to ensure that the water supplied is safe for the consumer [4]. Water pollution can cause adverse health effects for a representative number of people over predictable periods of time and is due to population growth, industrial development, and urbanization [5]. Unfortunately, clean, pure, and safe water only exists briefly in nature and is immediately polluted by prevailing environmental factors and human activities. Water from most sources is therefore unfit for immediate consumption without some sort of treatment [4].

The industrial pollutants associated with organic matter, inorganic dissolved solids, and other unwanted chemicals cause serious problems in the water quality [5]. Water-related diseases continue to be one of the major health problems globally due to the consumption of contaminated water. The high prevalence of diarrhea amongst children and infants can be traced to the consumption of unsafe water. The examination of microbiological river water quality according to technical standards is obligatory for use-related aspects such as drinking water production, irrigation, or recreation [6].

Water quality characteristics of aquatic environments arise from a multitude of physical, chemical, and biological interactions [8-10]. The public health significance of water quality cannot be overemphasized. Many infectious diseases are transmitted by water through the fecal-oral route [7]. Certain physical, chemical, and microbiological standards are designed to ensure the safety of water. It is on this premise that the research was carried out to assess the physiochemical and microbiological assessment of groundwater. In Ipogun in Ondo State, Nigeria.

Area of Study

This study was restricted to settlements in Ipogun (7°1853N: 5° 0448E) of Ondo State, Nigeria, with a population of about 176,327. Ipogun is a village opposite Ilara-Molkin; it lies about 208 miles (334 km) to the southwest of the capital, Abuja. The climate of Ipogun is that of the tropical rainforest type, with distinct wet and dry seasons. The annual rainfall varies from 2,000 mm to 1,150 mm. The mean monthly temperature is 27 degrees Celsius, while the mean relative humidity is over seventy-five percent. Schistosomiasis is endemic in the area. Another menacing ecological problem is the accelerated soil erosion.

Sampling

Six experimental sites were selected from the present study to analyze groundwater samples for various physiochemical and microbiological assessments.

Six samples were randomly collected, three each, from two settlements as follows: Settlement A comprised one well and one borehole water sample from the police station area in Ipogun. Settlement B comprised two wells and two borehole water samples collected from the Petroleum Trust Fund (PTF) national rural water supply program. All the samples were collected in sterile bottles following aseptic procedures and labelled accordingly. The samples were taken immediately to the microbiology laboratory at Elizade University, Ilara-Mokin, for analysis.

Method of Data Collection

Physicochemical Analysis of the Water Samples

The water samples were analyzed for pH, temperature, conductivity, hardness, acidity, and alkalinity.

pH

pH was determined by using a pH meter at the site of collection before transporting to the laboratory and was recorded [11].

Conductivity

Conductivity (EC) was determined by a conductivity meter. An electrode is connected to the meter, immersed into the sample of water so that the water will cover the sensitized electrode, and readings were recorded using an HANNA instrument [12].

Temperature

The temperature was obtained by using a HANNA pH meter, and readings were recorded for each water sample [13].

Hardness

50 ml of water sample was measured in a 250 cm conical flask, and 1 cm of ammonia buffer was added. 2-3 drops of Eriochrome Black T indicator were also added, and then the solution was titrated against 0.01 m EDTA solution until the wine-red color of the solution turned pale blue. Hardness is most commonly expressed as milligrams of calcium carbonate equivalent per liter. Water containing calcium carbonate at concentrations below 60 mg/l is generally considered soft; 60–120 mg/l is moderately hard; 120–180 mg/l is hard; and more than 180 mg/l is very hard [14].

Acidity

100 ml of a well-mixed sample was poured into a conical flask, 2-3 drops of phenolphthalein were added to the sample, and then it was titrated against 0.025 ml of NaOH till a pink color developed [15].

Alkalinity

50 ml of the water sample was measured into a conical flask, and 2-3 drops of bromcresol green were added and then titrated against 0.02N sulfuric acid until the color changed from blue to pale yellow [15].

Test for Metals

100 ml of each sample were measured into a beaker, and 5 ml of nitric acid was added to each sample so that the dissolved metals are kept in ionic form because this will enable the detection of the dissolved metals by the AAS machine. Then they were placed on a hot plate, and 20-25 ml were allowed to evaporate before removing them from the hot plate and allowing them to cool. Make the mark back to 100 ml by adding distilled water.

Materials Used

The materials used were a spirit lamp, gloves, cotton wool, ethanol, an inoculating loop, test tubes, test tube racks, foil paper, Petri dishes, a gram staining kit, conical flasks, an incubator, a refrigerator, a measuring cylinder, paper tape, a colony counter, a weighing balance, an autoclave, nutrient agar, Eosine methylene blue agar, Mueller Hilton agar, and water samples collected.

Microbiological Analysis of the Water Samples

Nutrient Agar Preparation: 6.72 g was weighed and diluted into 240 ml of distilled water in a 250 cm conical flask. The agar was prepared according to the manufacturer's specifications. The conical flasks were plunged with cotton wool and wrapped with foil to prevent contamination, and then the medium was autoclaved at 121°C for 15 minutes and allowed to cool to an extent (it was friendly to touch). 0.5 ml of each water sample was pipetted into the Petri dish aseptically, and the nutrient agar was poured into the Petri dish. The Petri dish was slightly moved in an anticlockwise direction to enable the agar that was poured into it to set and spread evenly. Having solidified, the medium was then incubated at 37°C for 24 hours.

Potato Dextrose Agar: 7.8 g was weighed and diluted into 200 ml of distilled water in a 250 cm conical flask. The agar was prepared according to the manufacturer's specifications. The conical flasks were plugged with cotton wool and wrapped with foil to prevent contamination, then the medium was autoclaved at 121°C for 15 minutes and allowed to cool to an extent (it was friendly to touch). 0.5 ml of each water sample was pipetted into the Petri dish aseptically, and the potato dextrose agar was poured into the Petri dish. The Petri dish was slightly moved in an anticlockwise direction to enable the agar that was poured into it to set and spread evenly. Having solidified, the medium was then kept at room temperature for 3 days.

Eosin Methylene Blue Agar: 7.2 g of EMB were weighed and diluted into 200 ml of distilled water in a 250 cm conical flask according to the manufacturer's specifications. The conical flasks were plunged with cotton wool and wrapped with foil to prevent contamination, and then the medium was autoclaved at 121°C for 15 minutes and allowed to cool to an extent (it was friendly to touch). 0.5 ml of each water sample was pipetted into the Petri dish aseptically, the medium was poured into the Petri dish, and the Petri dish was slightly moved in an anticlockwise direction to enable the agar that was poured into it to set and spread evenly. It was allowed to solidify before being incubated at 37°C for 24 hours.

Purification of Isolates

After incubation, colonies that developed on plates were randomly picked and subcultured until pure cultures were obtained on corresponding agar plates for purification and preserved on appropriate agar slants, kept in the refrigerator at 4-8ºC for further studies.

Identification of Isolates

Bacterial isolates were characterized to the generic level and, where possible, to the species level on the basis of their cultural features (i.e., color, shape, edge, elevation, etc.), morphological features (such as motility, gram reaction, cell arrangement, and shape), and biochemical features. The results obtained were then compared with standard references for proper identification of the isolates. Bergey’s manual was used for bacteria identification.

Morphological Characteristics

The morphological characteristics of the individual colonies that grew on the petri dishes were observed and recorded for their shape, size, margin, elevation, color, opacity, surface, and consistency. (Brooks et al., 2016).

Gram Staining

Gram staining was done to check if the microorganism present was gram positive or negative.

Catalyst Test

A smear was made on the glass slide with a drop of water and the microorganism. Few drops of hydrogen peroxide were added to the smear. Presence of bubbles indicates a positive sample and negative otherwise.

Indole Test 

Tryptophan broth was inoculated with broth culture, and the isolated colony of the test organism was emulsified in the broth. The setup was incubated at 37°C for 24–48 hours. 0.5 ml of Kovacs reagent was added to the broth culture. A color change of pink on top of the medium forming a ring shape indicated that the organism was present.

Motility Test

0.75 g of nutrient agar and 1.3 g of nutrient broth were measured and diluted into 100 ml of distilled water. 5.75 ml of the medium was dispensed into the test tube and autoclaved. The medium was allowed to cool and became semisolid before dipping inoculums straight into the test tube and incubating for 24 hours at 37°C.

Fermentation of Sugar

Different types of sugar were used, such as lactose, mannitol, arabinose, maltose, glucose, and fructose. 1 g of each sugar was weighed into different conical flasks and labeled accordingly. Into each flask, 1 g of peptone was added and made up to 100 ml with distilled water. 0.01 g of phenol red was added as an indicator. About 5 ml each of the 100 ml sugar solution was dispensed into different test tubes with Durham tubes inserted into each in an inverted form. The tubes were labeled appropriately with their mouths plugged with cotton wool and sterilized in an autoclave for 15 minutes at 121°C. These tubes were allowed to cool down before inoculation. Bacterial isolates were inoculated into the sugar solution inside the test tubes and incubated at 37°C for 72 hours. After incubation, the tubes were observed for acid production by a change in color from red to yellow. The change tubes were compared with the control to ascertain any color change; tubes were also examined for accumulation of space in the inverted Durham tubes. The presence of space indicates the production of gas as a result of utilization of the sugar by the inoculated organism, which brought about a color change [16].

Methyl Red Test

Each of the bacteria was aseptically inoculated into appropriately labelled and sterilized nutrient broth. Using a sterile inoculating loop, the tubes were incubated at 37°C for 24 hours. After incubation, 5 drops of methyl red indicator were added to the culture broth. The tubes were then examined for color change; the formation of red color is a positive test while the formation of yellow color is a negative test [17].

This procedure was repeated for all six water samples.

Statistical Analysis of Data

All data generated through analysis of water was analyzed statistically using mean and percentage.

The results of the data analysis for the research questions are presented below. The various levels of physiochemical parameters of well water and borehole water in Ipogun were shown in tables 4a and 4b. The average microbial load of water samples from Ipogun village is shown in table 5. Generally, the well water samples had higher bacterial loads than the borehole water samples. The highest bacterial load of 8.7×104 cfu/ml was obtained from the well 3 sample, while the lowest of 81.2×103 cfu/ml was obtained from borehole 1. A total of five (5) bacteria were identified from the water samples. They are Proteus vulgaris, Escherichia coli, Serratia marcescens, Bacillus cereus, and Pseudomonas aeruginosa, as shown in tables 3a and 3b, respectively. Only one Gram-positive bacterium was isolated from the water samples.

The fungi isolates from the water samples were subjected to both macroscopic and microscopic examination for identification. A total of seven (7) different fungi were isolated from the water samples. They are Aspergillus niger, Aspergillus flavus, Penicillium italicum, Rhizopus stolonifer, Mucor mucedo, Fusarium oxysporum, and Pithomyces species. The results of fungi isolation are presented in Table 1.

Research question one: What are the levels of physiochemical parameters of well water and borehole water in Ipogun?

Table 1: Comparison of Ground Water Quality with Drinking Water Standards.

Table 1 above shows the various levels of physiochemical parameters of well water and borehole water in Ipogun.

Table 2: Results of the Quantitative Amount of Some Heavy Metals in Water Samples.

Table 2 shows the quantitative amount of some heavy metals in the water samples.

pH

After observation of pH in the well and borehole water samples, settlement B had the highest pH (6.90) among all the study sites, and the lowest pH (6.20) was found at settlement A. However, in the case of well water, the highest pH (6.90) was found at well 2 and the lowest (6.20) at well 1. In the case of borehole water, the highest pH was found at BH3 and the lowest at WL1. The pH values obtained from all the samples ranged from 6.20 to 6.90. Which is low as compared to the standard limit. Therefore, all the pH values of the samples fell within the WHO permissible limits of 6-8.50.

Conductivity

The conductivity of groundwater from settlement A ranged from 50.5 to 75.5 µS/cm, and that of settlement B was 53.6 to 80.1 µS/cm. In all the samples analyzed, well water 2 had the highest conductivity while B.H1 had the least conductivity values. However, all the electrical conductivity values were within the WHO standard.

Alkalinity

The alkalinity (ALK) values of all the sampled water ranged from 6-15.4 mg/l. Alkalinity values from settlement A ranged from 6-10.5 mg/l, and settlement B alkalinity values ranged from 4-15.4 mg/l. All these values of alkalinity from the sample areas fell below the WHO standard limit of 200 mg/l.

Acidity

The acidity values of the sampled water ranged from 18.4 mg/l to 42.5 mg/l. Acidity values from settlement A ranged from 18.4 to 42.5 mg/l, and settlement B ranged from 24.1 to 37.5 mg/l. All these values of acidity from the sample areas fell below the standard limit because all pH levels were below 7.

Total Hardness

The range of total hardness (TH) for settlement A was 30-50 mg/L, while for settlement B it was 23.6-70 mg/L. These values are within the WHO standard of 60-180mg/l, below 60 mg/l = soft, 60-120 mg/l = moderately hard, 120-180 mg/l = hard, and more than 180 mg/l = very hard. (McGowan).

Cadmium

Cadmium (Cd) was analyzed in well and borehole water from all six selected sites. The highest cadmium concentration was observed in well water source WL3 (0.059 mg/l) at settlement B, and the lowest was at settlement A borehole source B.H1 (0.007 mg/l). However, all Cd values ranged from 0.007 to 0.059 mg/l. The amount of Cd present in BH2, B.H3, WL1, WL2, and WL3 was above the permitted WHO standard.

Lead

The amount of lead (Pb) ranged from 0.20 to 0.52 mg/l. Settlement A showed a range of 0.26-0.52 mg/l, while settlement B values ranged from 0.20-0.49 mg/l. The highest concentration of Pb was observed in WL1 (0.52 mg/l) from settlement A, and the lowest concentration of Pb was observed in WL2 from settlement B. The amount of Pb present in B.H1, B.H3, WL2, and WL3 was below the WHO standard.

Zinc

The amount of zinc (Zn) ranged from 0.0042 to 0.0390 mg/l. Settlement A showed a range of 0.0090-0.0465 mg/l, while settlement B values ranged from 0.0042-0.0390 mg/l. The highest concentration of Zn was observed in BH1 (0.0465 mg/l) from settlement A, and the lowest concentration of Zn was observed in WL3 (0.0042 mg/l) from settlement B. The amount of Zn present in the samples was within the WHO standard.

Iron

The amount of iron in the samples ranged from 0.072 to 0.713 mg/l. Settlement A showed a range of iron values from 0.072-0.351 mg/l, while settlement B gave values for iron ranging from 0.095-0.713 mg/l. However, all the values of iron were within the WHO standards of 0.30 mg/L.

Copper

Copper (Cu) was not detected (ND) in all the water samples.

Research Question Two: What are the levels of bacteriological parameters of well water and borehole water in Ipogun?

Table 3: Average Microbial Load from the Water Samples.

Table 3 above shows the various levels of Bacteriological parameters of well water and borehole water in Ipogun.

Table 4: Colonial Morphology of Bacteria Isolated from Water Samples.

Table 5: Biochemical and Sugar Fermentation Reactions of Isolates.

Keys: Cat = catalase, Oxi = oxidase, Ind= indole, H₂S = Hydrogen Sulphide, Nit red = nitrogen reduction, Ure = urease, Lact = lactose, Fruc = fructose, Malt = maltose, Gala = galactose, Glu = glucose, Arab = arabinose, Raf = raffinose, Man = mannitol, MR = methyl red, VP = Voges-Proskauer; - = negative; + = positive.

Table 6: Macroscopic and Microscopic Identification of Isolated Fungi from the Water Samples.

Discussion of Results

The Levels of Physiochemical Parameters of Borehole and Well Water in Ipogun

The pH values indicate that the groundwater of the sample areas is low and slightly acidic. Low pH of surface water of various rivers and other water sources may be due to the dilution effect of rainwater during the rainy season, as reported by various workers. pH controls the chemical state of many nutrients, including dissolved oxygen, phosphate, nitrate, etc. Due to this effect, aquatic organisms are affected by pH changes as their metabolic activities are pH dependent. Nonetheless, the pH has no direct adverse effects on health. According to Adefemi and Awokunmi [18], acidic water results in corrosion of iron and steel materials such as pipes, clogging of distribution pipes causes objectionable taste of drinks and food, and stains clothes and rusts cooking utensils. Acidity of bottled water has been reported by Wright [19].  However, higher values of pH hasten the scale formation in water heating apparatus and also reduce the germicidal potential of chloride, resulting in the formation of trihalomethanes, which are toxic in nature [20].

Electrical conductivity (EC) is a measure of the ability of water to conduct electrical current. This is measured in terms of the amount of ions in a solution. The more the ions in the solution, the higher is the conductivity. Based on the fact that seawater has higher conductivity in comparison to fresh water. Thus, it could be concluded that conductivity in water is affected by the presence of inorganic dissolved solids such as chloride, nitrate, sulfate, and phosphate anions or sodium, magnesium, calcium, iron, and aluminum cations. Various phenomena are reported in the literature that cause enhancement in the value of EC, such as natural enrichment in electrolytes, phenomena of mineralization, or weathering of sediments [21].

Alkalinity of water is defined as the ionic concentration that can neutralize the hydrogen ions. The phenolphthalein alkalinity value is zero, indicating the absence of any carbonate and hydroxyl ions. The bicarbonate alkalinity is expressed as a total alkalinity that ranges between 4-15 mg/l. The alkalinity values of all samples of the present study are below the permissible limit of 20-200 mg/l [22].

The WHO international standard for drinking water classified water with a total hardness of CaCO₃ as <60 mg/l as soft, 60-120 mg/l as moderately hard, 120-180 mg/l as hard, and more than 180 mg/l as very hard. Based on this classification, the water samples analyzed from BH1, BH2, BH3, and WL1 are soft, and water samples from WL2 and WL3 are moderately hard. Thus, the waters are suitable for domestic use in terms of hardness. This is because moderately hard water is preferred to soft water for drinking purposes, as hard water is associated with a low death rate from heart diseases [18].

The major sources of cadmium in drinking water are corrosion of galvanized pipes, erosion of natural deposits, discharge from metal refineries, and runoff from waste batteries and paints. Once cadmium is in the air, it spreads with the wind and settles onto the ground or surface water as dust.Cadmium has the chronic potential to cause kidney, liver, bone, and blood damage from long-term exposure at levels above the MCL. From the analysis of all six water samples, it was observed that the amount of cadmium present in the water samples (wells and boreholes on Ipogun) is above the permissible WHO standard. 

Lead (Pb) leaches into water through corrosion—a dissolving or wearing a way of metal caused by a chemical reaction between water and your plumbing. The result from analyzing all six samples from settlements A and B in Ipogun shows that only the water sample from BH2 in settlement B is within the WHO standard, while water samples from BH1, BH3, WL1, WL2, and WL3 contain amounts of Pb below the acceptable WHO standard. Lead is harmful to children and adults. Adults exposed to lead can suffer from cardiovascular effects, increased blood pressure, and increased incidence of hypertension. Decreased kidney function.

High natural levels of zinc in water are usually associated with higher concentrations of other metals such as lead and cadmium. The concentration of Zn in all six samples was observed to be below the permissible limit of the WHO standard (0.35-1.19 mg/l) because they ranged from 0.0090-0.0465 mg/l.

From the result gotten after analyzing all six water samples for iron (Fe), it was observed that the amount of Fe present in the samples is below the permissible limit of the WHO standard. This can lead to clogging of pipes, can leave brownish stains on laundry, reddish-brown particles on fixtures that are hard to remove, and can cause an unpleasant taste and odor in water.

The Levels of Microbiological Parameters of Borehole and Well Water in Ipogun

The high bacteria pollution observed in the study may be attributed to both the shallow depth at which water is tapped, settlement patterns, and land use practices. The location of groundwater also does not take water quality into cognizance. Wells and boreholes are often located too close to sanitation systems, and almost all the wells used bailers as a mode of collection. The bacterial pollution of shallow wells around Ipogun is anthropogenic in origin. The high human concentration in these locations enhances the use of pit latrines and septic soakaways, which are often located too close to the boreholes and wells in most households. In addition to this, free-ranging domestic animals and other domestic solid wastes that are dumped around the houses are possible sources of bacteria pollution of the shallow wells. Besides, there is a tendency for a higher rate of infiltration of precipitated water during the rainy season. A high level of fecal bacteria in water samples indicates the possible presence of pathogenic (disease-causing) organisms [23]. From the result of the bacteria identification carried out, it is evident that diseases such as cholera, typhoid fever, bacterial dysentery, infectious hepatitis, and food poisoning can possibly result from the consumption of the untreated water. Eye, ear, nose, and throat infections can also spread from contact with the water.

In conclusion, the pH level of the water sample (well and borehole) from all sites was below 7, which led to the low acidity of each sample. Hence, all water samples were slightly acidic in nature. Metals such as cadmium and lead levels were high, and continuous intake of the water can lead to health risks. However, the levels of bacteria suggest there is a high incidence of contamination of well and borehole water by the pathogenic organisms and the presence of some opportunistic microorganisms [23]. To reduce the widespread incidence of contamination of borehole and well water, it is advocated that wells dug must be deep and covered adequately. Subsequently, there is a need to establish sewage treatment plants in major human settlements so that untreated sewage would not contaminate boreholes.

Recommendations

1. Regular and quantified monitoring of geochemical characteristics of ground water for sustainable water management.
2. Boiling and other disinfection methods of borehole and well water should be done before usage of the water.
3. Deep wells should be constructed and adequately covered to prevent contamination.
4. Pit latrines and septic tanks should be located very far away from boreholes and wells.
5. Good sanitary condition of wells should be maintained at all times to minimize the contamination of the well water.
6. Provision of clean, reliable and portable water can be provided to the people residing in Ipogun by the Government.
7. Elimination of cadmium sources from the environment of Ipogun.

1. Taiwo AG, Adewunmi AR, Ajayi JO, Oseni OA, Lanre-lyanda YA. Physico-Chemical and Microbial Analysis of the Impact of Abatoir Effluents on Ogun River Course. Int J Chem Tech Res. 2014; 6: 3083-3090.
2. Rajiv P, Hasna AS, Kamaraj M, Rajeshwari S, Sankar A. Physico Chemical and Microbial Analysis of Different River Waters in Western Tamil Nadu, India. Int Res J Environ Sci. 2012; 1: 2-6.
3. Karikari AY, Ansa-Asare OD. Physico-Chemical and Microbial Water Quality Assessment of the Densu River of Ghana. West Afr J Appl Ecol. 2006; 10: 87-100.
4. Omezuruike OI, Damilola AO, Adeola OT, Fajobi EA, Shittu OB. Microbiological and Physicochemical Analysis of Different Water Samples Used For Domestic Purposes in Abeokuta and Ojota, Lagos State, Nigeria. African Journal of Biotechnology. 2008; 7: 617-621.
5. Radha R, Dharmaraj K, Ranjitha BD. A Comparative Study on the Physicochemical and Bacterial Analysis of Drinking, Borewell and Sewage Water in the Three Different Places of Sivakasi. J Environ Biol. 2007; 28: 105-108.
6. Kavka GG, Kasimir GD, Farnleitner AH. Microbiological Water Quality of the River Danube (Km 2581-Km 15): Longitudinal Variation of Pollution as Determined by Standar Parameters. Proceedings of the 36th International Conference of IAD, Austrian Committee Danube Research/IAD, Vienna. 2006; 415-421.
7. Shittu OB, Olaitan JO, Amusa TS. Physico-Chemical and Bacteriological Analyses of Water Used for Drinking and Swimming Purposes in Abeokuta, Nigeria. Afr J Biomed Res. 2008; 11: 285-290.
8. Abd Al-Kareem AF, Al-Arajy KH, Jassim KA. Microbiological Analysis on Tigris River Water in the Selected Sites in Baghdad Province, Iraq. J Environ Earth Sci. 2015; 5: 60-65.
9. Asbolt NJ. Microbial contamination of drinking water disease out comes in developing Toxicology. 2004; 198: 229-238.
10. Asbolt NJ, Grabow WOK, Snozzi M. Indicators of microbial water quality. In: Fewrell, L.J. bartram (Eds), water Quality: Guidelines Standards and Health. 2001.
11. Baghel VS, Gopal K, Dwivedi S, Tripathi RD. Bacterial Indicators of Faecal Contamination of the Gangetic River System Right at Its Source. Ecol Indic. 2005; 5: 49-56.
12. Baird C, Cann M. Environmental Chemistry (3rd edition). W.H. Freeman, USA. 2004.
13. Carpenter SR, Caraco NF, Correll DL, Howarth RW, Sharpley AN, et al. Nonpoint pollution of surface waters with phosphorus and nitrogen. Ecol Appl. 1998; 8: 559-568.
14. Dan’Azumi S, Bichi MH. Industrial pollution and heavy metals profile of Challawa River in Kano, Nigeria. Journal of Applied Sciences in Environmental Sanitation. 2010; 5: 23-29.
15. Jain CK, Kumar S, Rao YR. Trace element contamination in a coastal aquifer of Andhra Pradesh. Pollution Research. 2004; 23: 13-23.
16. Kazi TG, Arain MB, Jamali MK, Jalbani N, Afridi HI, et al. Assessment of water quality of polluted lake using multivariate statistical techniques: A case study. Ecotoxicol Environ Saf. 2009; 72: 301-309.
17. Ladokun OA, Oni SO. Physico-Chemical and Microbiological Analysis of Potable Water in Jericho and Molete Areas of Ibadan Metropolis. Adv Biol Chem. 2015; 5: 197-202.
18. Guidelines for drinking water quality, 4^th edition. Recommendation 1, Geneva: World Health Organization (WHO). 1996.
19. Wright KF. Is your drinking water acidic? A comparison of the varied pH of popular Bottled waters. American Dental Hygenists Association. 2015; 89: 6-12
20. Kumar A, Bisht BS, Talwar A, Chandel D. Physico-chemical and microbial analysis of ground water from different regions of Doon valley. Int J Appl Env Sci. 2010; 5: 433-440.
21. Abegaz SM. Investigation of input and distribution of polluting elements in TinishuAkaki River, Ethiopia, based on the determination by ICP-MS. PhD Thesis, Ghent University, Belgium. 2005; 221.
22. Tripathi DK, Pandey G, Jain CK. Physicochemical Analysis of Selected Springs water samples of Dehradun city, Uttarakhand, India. Int J Innov Res Sci Technol. 2015; 2: 99-103.
23. Chris W. The safe drinking water Act: A blueprint for protecting the nation’s drinking water. Water quality and health council 2020. 2014.