ANALYSIS OF THE PHYSICO-CHEMICAL
AND BACTERIOLOGICAL PARAMETERS OF BOTTLED WATER
AVAILABLE IN KATHMANDU VALLEY

A Case Stu...
LETTER OF RECOMMENDATION
This is to certify that Ms. Ranjana Budhathoki, a student of M.Sc. 1st year, Central
Department o...
ACKNOWLEDGEMENT

This case study entitled “Analysis of Physico-chemical and Bacteriological Parameters of
Bottled Water Av...
ABSTRACT
Water is vital for life. However, it also serves as the commonest route of transmission of a
number of infectious...
ACRONYMS
ADB

Asian Development Bank

APHA

American Public Health Association

BMC

Beverage Marketing Corporation

CBS

...
TABLE OF CONTENTS

Letter of Recommendation
Acknowledgement
Abstract
Acronyms
Table of Contents
Lists of Tables
List of Fi...
LIST OF TABLES
Table 1.1.5.1: Guidelines of Drinking Water Quality (Source: FAR
(2062), WHO (1994), EPA (2006)) and Nepal ...
CHAPTER-1
INTRODUCTION

1.1. BACKGROUND:
Water is inevitably essential to sustain life. Out of total 3% of fresh water in ...
The quality of water is reflected by various physical, chemical and biological conditions
which in turn are influenced by ...
inadequate sanitation, pollution or unavailability of water. Hence it is necessary to purify
and disinfect water before it...
independent testing done for bottled water, problems are periodically discovered. Many
individual researches and studies i...
Limited (KUKL) produces about 120 million liters per day (MLD) in wet season and 80
MLD in dry season whereas the demand i...
of brands of bottled water including jar water are available in the market at different price
rates and therefore, doubt a...
1.2. STATEMENT OF THE PROBLEM:
Water quality has a direct impact on public health. More than 80% of deaths is caused due
t...
CHAPTER- 2
LITERATURE REVIEW

Sharma S., (1978) analyzed the household drinking water in 39 localities of Kathmandu
valley...
bottles (17.4%) and 5 out of 23 bottles were found to have less content, i.e., water had
leaked while transportation. The ...
techniques in a bottled water company. He analyzed water from water tank, water filter
and bottled water twice monthly for...
CHAPTER- 3
METHODOLOGY
3.1. SAMPLE COLLECTION, TRANSPORTATION AND PROCESSING
Samples of nine brands of jar water of 20 lit...
Table 3.3.1.: Methodology of Physico-chemical parameters and microbial analysis
S. No.
1
2
3
4
5
6
7
8
9
10
11
12
13

Para...
Calculation
Chloride (mg/L) = (a-b) × N ×35.5 × 1000
V
Where, a = Volume of titrant (silver nitrate) for sample
b= Volume ...
Calculation
Free CO2 (mg/L) = A × Normality of NaOH × 44 ×1000
Volume of sample in mL

Where, A = Volume of NaOH used in m...
Total Alkalinity (mg/L) = a ×N×1000×50
mL of sample

where, a= Volume of standard H2SO4 consumed in titration
N= Normality...
14. Total Iron
Iron content in the water sample was determined by colorimetric method. In this method
50 mL of the sample ...
negative result were further incubated at 37° C for 24 hours. The tubes showing gas
formation and acid formation were furt...
CHAPTER- 4
RESULTS AND DISCUSSION

The table with respective values of each of the parameters is shown in the annex.

1. p...
2. Temperature:

Fig 4.2: Graph showing temperature values

Temperature of fresh water varies normally from 0 to 35ºC depe...
Electrical conductivity is the measure of the capacity of water to conduct electric current.
The values of conductance ran...
5. Hardness:

Fig 4.5: Comparison between the hardness values

Hardness is imparted to the water mainly by calcium and mag...
6. Free CO2:

Fig 4.6: Graph showing values of Free CO2 in two tests
Surface water normally contains less than 10mg/L of f...
Chloride is present in appreciable amounts in all natural water. Concentration varies from
few milligrams to several thous...
9. Total Iron:

Fig 4.9: Graph showing the values of Total Iron

Iron has got a little concern as health hazard, but it st...
according to WHO. The values obtained are in the range of 0.01-0.02 mg/L in both tests,
all below the permissible limits.
...
13. Bacteriological examination:
Table 4.13.1 Bacteriological examination water during January/February
Brands

Presumptiv...
In this test, only one brand was found to be devoid of both total and fecal coliform while
two other brands showed total c...
was obtained for brand 4. The values of DO ranged from 6.9 – 9.3 mg/L. The value of
iron was obtained in a range of 0.05-0...
CHAPTER- 5
CONCLUSION AND RECOMMENDATION

5.1. CONCLUSION:
Hence from the above results, it can be concluded that jar wate...
5.2. RECOMMENDATIONS:
1. There are varieties of jar water and their quality also varies. Thus it is necessary to pick
up t...
REFERENCES
ADB(2004) Water for all: The impact of water on the poor, Asian Development Bank,
Manila.
APHA. ,1998: Standard...
Pandey B., (2009), “A Case Study of Drinking Water Quality Status in Central
Development Region, Nepal”

Pimentel D., Berg...
ANNEX

Results of the Physio-chemical tests:
Table- Physio-chemical parameters of jar water during January/February

Sampl...
Table- Physio-chemical parameters of jar water during February/March

Sample(B)

Brand Brand Brand Brand Brand
5

Brand Br...
MPN DETERMINATION FROM MULTIPLE TUBE TEST
95
PERCENT
NUMBER OF TUBES GIVING
MFN
CONFIDENCE LIMITS
POSITIVE REACTION OUT OF...
Composition of Eosin Methylene Blue (EMB)
Peptic digest of animal tissue
10.00 gm / Ll
Dipotassium phosphate
2.00 gm / L
L...
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Case study of_jar_water_in_kathmandu_valley-ranjana_budhathoki (1) BY Muhammad Fahad Ansari 12IEEM14

  1. 1. ANALYSIS OF THE PHYSICO-CHEMICAL AND BACTERIOLOGICAL PARAMETERS OF BOTTLED WATER AVAILABLE IN KATHMANDU VALLEY A Case Study on the Partial Fulfillment of the Requirements for M.Sc., First Year, Environment Science, T.U. Submitted To: Central Department of Environment Science Tribhuwan University Kirtipur Submitted By: Ranjana Budhathoki M.Sc. 1st year Group A Symbol number- 4374 August, 2010
  2. 2. LETTER OF RECOMMENDATION This is to certify that Ms. Ranjana Budhathoki, a student of M.Sc. 1st year, Central Department of Environment Science, T.U., has carried out this case study entitled “Analysis of Physico-chemical and Bacteriological Parameters of Bottled Water Available in Kathmandu Valley” under my supervision and guidance. She is a sincere student and has performed all the works in field as well as laboratory with full dedication and satisfaction. So, I recommend this case study report for final approval. _________________________ Ms. Ushana Shrestha (Supervisor) Central Department of Environmental Science Tribhuvan University II
  3. 3. ACKNOWLEDGEMENT This case study entitled “Analysis of Physico-chemical and Bacteriological Parameters of Bottled Water Available in Kathmandu Valley” is a product of research, knowledge and support of many individuals as well as organizations to whom I would like to express my sincere gratitude. I am truly indebted to my supervisor Ms. Ushana Shrestha for her supervision, guidance and invaluable suggestion. Her recommendations and suggestions have been a key for the successful completion of my case study. I would also like to express my gratitude to the concerned officials of Company Registrar’s Office and Kathmandu Upatyaka Khanepani Limited (KUKL) for providing necessary information. I am very much thankful to my classmates for their constant support, co-operation and motivation throughout the study. I would also like to thank the lab staff for their ever ready assistance. I would like to thank all other respected teachers of Central Department of Environmental Science, Tribhuvan University, Kirtipur for their continuous aspiration and motivation. Ranjana Budhathoki III
  4. 4. ABSTRACT Water is vital for life. However, it also serves as the commonest route of transmission of a number of infectious diseases. The WHO has estimated that up to 80% of all sickness and disease in the world is caused by inadequate sanitation and polluted water. Nepal faces a number of problems regarding both its drinking water quality and availability. The municipal water supplies are inconsistent and unreliable. Not only the shortages in quantity, but also the compromised quality of municipal tap water has become a major public health issue (Warner N.R. et al., 2007). Throughout Nepal, people are facing health problems resulting from water contamination. In the context of growing health consciousness and chronic water shortages, most of the urban residents have switched to bottled water as a safe alternative. The public perception is that bottled water is regularly of high quality. This belief is encouraged by publicly reported problem of municipal tap water as well as the public perception of purity driven by advertisements. However, many studies have shown that these beliefs need not always be true. Samples of nine brands of jar water of 20 ml capacity were analyzed twice for various physicochemical as well as bacteriological parameters during January, February and March. All the physicochemical parameters like pH, DO, hardness, alkalinity, chloride were within WHO acceptable limits. Ammonia was detected in some samples but is within the limits set by WHO. Iron and Nitrates were found in small quantities but within the limits set by WHO. From the bacteriological point of view, 66% of the total samples were heavily contaminated with coliforms during the test in January/February. During the test in February/March, 89% of the total sample was found to be contaminated with total coliforms whereas 66% were contaminated with fecal coliforms. From the analysis of jar water marketed in Kathmandu valley, it was concluded that the jar water samples are heavily contaminated with coliform bacteria and unsatisfactory for drinking purpose. IV
  5. 5. ACRONYMS ADB Asian Development Bank APHA American Public Health Association BMC Beverage Marketing Corporation CBS Central Bureau of Statistics CFU Colony Forming Units DISVI Italian International Co-operation EDTA Ethylene Diamine Tetra Acetic Acid EMB Eosin Methylene Blue ENPHO Environment and Public Health Organization EPA Environment Protection Act FAR Food Act Regulation ICIMOD International Center for Integrated Development JICA Japan International Co-operation Agency KUKL Kathmandu Upatyaka Khanepani Limited MLD Million Liters per day MPN Most Probable Number NDWQS Nepal Drinking Water Quality Standards NRDC National Resources Defense Council PET Poly-Ethylene Terephthalate pH Percentage of Hydrogen Ion Concentration UN United Nations UNICEF United Nation Children Fund WHO World Health Organization V Mountain
  6. 6. TABLE OF CONTENTS Letter of Recommendation Acknowledgement Abstract Acronyms Table of Contents Lists of Tables List of Figures I II III IV V VI VI Chapter 1: Introduction 1.1. Background 1.1.1. Fresh Water Shortage 1.1.2. Water Quality 1.1.3. Bottled Water 1.1.4. Water Related Problems in Nepal 1.1.5. Bottled Water in Nepal 1.2. Statement of the Problem 1.3. Justification 1.4. Objectives 1.4.1. Broad Objective 1.4.2. Specific Objectives Chapter 2: Literature Review Chapter 3: Methodology 3.1. Sample collection, transportation and processing 3.2. Sampling frequency 3.3. Analysis of water samples 3.3.1. Analysis of physicochemical parameters of water samples 3.3.2. Analysis of microbial variables of water sample Chapter 4: Results and Discussion Chapter 5: Conclusion and Recommendations 5.1. Conclusion 5.2. Recommendations 1 1 1 1 3 4 5 7 7 7 References Annex 31 33 VI 8 11 11 11 11 18 29 29 30
  7. 7. LIST OF TABLES Table 1.1.5.1: Guidelines of Drinking Water Quality (Source: FAR (2062), WHO (1994), EPA (2006)) and Nepal Drinking Water Quality Standards (2006) 6 Table 3.3.1: Methodology of Physio-chemical parameters and microbial analysis 12 Table 4.2.1: Results of the Presumptive Count during Jan/Feb (A) 26 Table 4.2.2: Results of MPN test during Feb/Mar (B) 26 LIST OF FIGURES Figure 4.1: Comparison between pH values 18 Figure 4.2: Graph showing Temperature values 19 Figure 4.3: Comparison of Conductance values in two tests 19 Figure 4.4: Comparison of Alkalinity values in two tests 20 Figure 4.5: Comparison between Hardness values 21 Figure 4.6: Graph showing the values of Free CO2 in two tests 22 Figure 4.7: Graph showing the values of Chloride in two tests 22 Figure 4.8: Comparison between DO values 23 Figure 4.9: Graph showing the values of Total Iron in two tests 24 Figure 4.10: Comparison between Nitrate-N values 24 Figure 4.11: Graph showing the values of Phosphate-P 25 Figure 4.12: Comparison between Ammonia-N values 25 VII
  8. 8. CHAPTER-1 INTRODUCTION 1.1. BACKGROUND: Water is inevitably essential to sustain life. Out of total 3% of fresh water in the earth, 77% are captured in the glaciers, 22% underground, 0.33% lakes, 0.18% soil moisture, 0.03% rivers and 0.03% in the atmosphere. Majority of freshwater are locked as glacier and polar ice which is difficult to utilize and importing them is costly. As fresh water resources are further stretched to meet the demands of industry, agriculture and an ever expanding population, the shortage of safe and accessible drinking water is estimated to become the major challenge in many parts of the world. There are two major problems with the fresh water of the earth: 1.1.1.. Fresh water shortage: Large volume of water is captured in the ocean as salt water. Since majority of the fresh water is stored in the glaciers and polar ice caps, there is a relative shortage of fresh water. Lakes and rivers are the primary sources of fresh water for human consumption but they only contain 0.26% of fresh water reserves and most of that is used for agriculture and industry (Shiklomanov, 1993; Gleick, 1993; Chapagain and Hoekstra, 2004). The remaining 30% stored as ground water, provides approximately 23% of the water used for the human consumption (Pimentel et al., 2004). In addition, the available fresh water is not uniformly distributed. 41% of the world’s population lives in areas characterized by either water stress or water scarcity (Global Environment Facility, 2002). Approximately, 1.1 billion people face chronic shortages of safe water for drinking and sanitation (United Nations, 2003; WHO/UNICEF, 2000).The condition is even worse in developing countries. In fact, 30% of the rural populations in many developing countries still obtain water from rivers dug pits, and other unsanitary sources (Olmstead, 2003). 1.1.2. Water Quality: Although water is vital for life, it also serves as the commonest route of transmission of a number of infectious diseases. Thus, water quality must be ensured before drinking and the water we drink must be safe. Safe drinking water is defined as water with microbial, chemical and physical characteristics that meet WHO guidelines of national standards on drinking water quality (WHO, 2007). 1
  9. 9. The quality of water is reflected by various physical, chemical and biological conditions which in turn are influenced by natural and anthropogenic sources. (ADB/ICIMOD).Water quality parameters like alkalinity, hardness, Dissolved Oxygen (DO), chloride, Total Dissolved Solid (TDS) etc add to the aesthetic value of water, while parameters like ammonia, lead, arsenic, nitrate etc may cause adverse health effects. Water having high or low pH, greater extent of turbidity etc. is objectionable to use. Appropriate amount of chloride content and hardness are desirable but higher content of the same makes the water unaesthetic. Similarly higher content of phosphate, nitrate, ammonia, iron, are undesirable. Some other chemical constituents like arsenic, lead etc. may be toxic. From microbiological point of view, drinking water should be free from any kinds of pathogens as well as opportunistic microflora. Although there are a number of microorganisms present in water that may pose health threat like Salmonella spp, Shigella spp, Coliforms, Mycobacterium spp etc., coliforms are used to assess water quality. Coliforms are gram negative rod shaped bacteria capable of growth in presence of bile salts and able to ferment lactose at 35-370 C with the production of acid, gas and aldehyde within 24-48 hours. They are oxidase negative and non-spore forming. Coliform organisms (E. coli) have long been recognized as a suitable microbial indicator of drinking water quality largely because it is easy to isolate and enumerate them. They are present in the intestine of warm blooded animals including humans. Thus, their presence in water samples indicates the presence of fecal matter and the possible presence of pathogenic organisms of human origin. If other pathogenic microbes are used as an indicator, then there is high chance of getting contaminated oneself and the sample taken should be large to trap pathogens, since their numbers decrease as they mix with water. The micro-organisms in water are capable of causing various diseases like typhoid, cholera, diarrhea, dysentery, hepatitis etc. According to WHO (2002), unsafe water supply is a major problem and fecal contamination of water sources and treated water is a persistent problem worldwide. Globally, 1.1 billion people rely on unsafe drinking water sources from lakes, rivers and open wells. The majority of these are in Asia (20%) and Sub-Saharan Africa (42%) (WHO/ UNICEF, 2000; WHO/ UNICEF-JMP, 2004). The use of these unsanitary sources helps to explain why 90% of human infections in less developed countries are caused by water borne diseases (Pimentel et al., 2004). The WHO has estimated that up to 80% of all sickness and disease in the world is caused by 2
  10. 10. inadequate sanitation, pollution or unavailability of water. Hence it is necessary to purify and disinfect water before it is available for drinking. Many researches and studies have revealed that tap water do not ensure the quality of water. According to the National Water Quality Association, 56% of all people are worried about the quality of municipally treated tap water. With the rising concern on public health, people choose bottled water over tap water. 1.1.3. Bottled Water: Bottled water is a term referring water that is presumed to be processed, packaged and sold in containers or simply bottles. According to the International Bottled Water Association, “Bottled water is a great beverage choice for hydration and refreshment because of its consistent safety, quality, good taste and convenience”. Bottled water can be categorized into Artesian well water, distilled water, mineral water, purified water, sparkling water, well water etc according to their source and state of purification. The mass production and marketing of bottled water exploded in the late 20th century. In the global scenario, sales and consumption of bottled water have skyrocketed in recent years. From 1988 to 2002, the sales of bottled water globally have more than quadrupled to over 131 million cubic meters annually (BMC, 2003). Bottled water sales worldwide are increasing at a rate of 10% annually (Bottled Water Web, 2003). One of the main reasons of this is the compromised water quality provided by municipality. Another reason may be the public perception that the bottled water is essentially of high quality. The public perception and probably the reality is that bottled water is regularly of high quality. This belief is encouraged by publicly reported problem of municipal tap water as well as the public perception of purity driven by advertisements and packaging labels featuring pristine glaciers and crystal clear mountain springs. However, many studies have shown that these beliefs need not always be true. A four-year study conducted by the National Resources Defense Council (NRDC, 1999) revealed that about one-third of the samples contained significant contamination, including synthetic chemicals, bacteria and arsenic, in at least one sample, out of more than 1000 samples of 103 bottled water brands tested. It also concluded that “an estimated 25% or more of the bottled water is really just tap water in bottle- sometimes further treated, sometimes not”. Even with limited 3
  11. 11. independent testing done for bottled water, problems are periodically discovered. Many individual researches and studies in developed countries have shown that only because water comes out of a bottle doesn’t mean that it is definitely purer and safer than the tap water. Similarly, in the words of NRDC, “While much tap water is indeed risky, having compared the available data, we conclude that there is no assurance that bottled water is any safer”. Environmental Effects of Bottled Water: 1. According to World Wide Fund for Nature (WWF), 2001 report, roughly 1.5 million tons of plastics are expended in the bottling of 89 billion liters of water each year. 2. The recycling rate of these bottles is minimum which produces a serious problem of waste management. 60 millions plastic bottles a day are disposed off in USA alone. 3. Energy required to manufacture and to transport these bottles to market severely drains limited fossil fuels. 4. Reusing plastic PET bottles compromises the water quality. Moreover there are ongoing arguments on the leaching of harmful chemicals from Poly-Ethylene Terephthalate (PET) bottles. 5. Bottled water industry is an exceptionally wasteful industry. More than three times water is needed to fill 1 bottle of water. 6. There may be local effects due to bottling plants. Also, the question for the sustainability of the bottled water sources may arise. In places where bottled water are filled from underground aquifers, they may get depleted over time. 1.1.4. Water Related Problems in Nepal: Nepal is a country with rich water resources. There are more than 6000 river and rivulets. The average annual runoff within the Nepalese territory is estimated at about 174 billion cubic meters. However, the management of this resource is very poor due to which many cities and towns of this country are facing severe shortages. Nepal faces a number of problems regarding both its drinking water quality and availability. Kathmandu, the capital of Nepal, suffers a severe drinking water supply crisis, which becomes more pronounced in the dry seasons. The existing water supply system of Kathmandu Upatyaka Khanepani 4
  12. 12. Limited (KUKL) produces about 120 million liters per day (MLD) in wet season and 80 MLD in dry season whereas the demand is around 320MLD. The KUKL supplies 40% of its water from surface water sources while the rest 60% comes from the underground sources. The municipal water supplies are inconsistent and unreliable. Not only the shortages in quantity, but also the compromised quality of municipal tap water has become a major public health issue. Throughout Nepal, people are exposed to severe health threats resulting from water contamination by sewage, agriculture and industry. Owing to the impact of sewage, typhoid, dysentery, and cholera are endemic every summer (Khadka, 1993). These diseases account for 15% of all illness and 80% of total deaths, but those number increases to 41% of all illness and 32% of all deaths in children up to 4 years (Sharma, 1990). Diarrhoeal diseases are recorded as the second most prevalent disease in Nepal. According to Sharma, 2003, around 75 children die each day from diarrhea alone. Recently, the diarrhoeal epidemic that affected the hills of mid and far-western districts like Jajarkot, Rukum, Dang, Humla, etc in Bhadra, 2066 taking the lives of more than 200 people also makes us more concerned about the drinking water quality. Although being the dwellers of the capital city, we are neither in the state to proclaim proudly that the water we use is any purer nor can it be guaranteed that these kinds of epidemics can’t occur in the valley. Thus, conveying message to the public about water quality and sanitation and at the same time, using disinfectants to purify water is a must in the present scenario. In the context of growing health consciousness and chronic water shortages, most of the urban residents have switched to bottled water as a safe alternative. 1.1.5. Bottled Water In Nepal: The history of bottled water in Nepal can be dated before 1992 when Star Water was the only brand available. Subsequently, Aqua Minerals Nepal Private Limited was established. Since then, the numbers of bottled water companies have been increasing. According to the recent data provided by Company Registrar’s Office, there are 55 companies of bottled water registered in the country out of which only few have received NS Standards. PET bottles of 1 liter and jars of 20 liters are available in the market. The price of PET bottles of 1 liter ranges from Rs.15-30 whereas for jar water ranges from Rs.100-750 with refilling price ranging from Rs.45-150. PET bottles are discarded after use while jars are taken back to the related companies and refilled. Jar water are commonly used for household purposes as well as in offices, educational institutions and restaurants. Varieties 5
  13. 13. of brands of bottled water including jar water are available in the market at different price rates and therefore, doubt about the quality can arise. The regulation and monitoring of the quality of bottled water is not proper. In Nepal, the drinking water quality is assessed with reference to the National Drinking Water Quality Standards (NDWQS). In addition to this, WHO and EPA guidelines are also followed. The respective guideline values are tabulated below: Table 1.1.5.1.: Guidelines of Drinking Water Quality (Source: FAR (2062), WHO (1994) EPA (2006)) and Nepal Drinking Water Quality Standards (2006): Parameters FAR WHO EPA NDWQS pH 6.5-8 6.5-8.5 6.5-8.5 6.5-8.5 TDS ( mg/L) 750 600 500 Alkalinity( mg/L) 600 - - Chloride( mg/L) 200 250 250 Total 200 200 - 500 DO( mg/L) - >5 - - Ammonia( mg/L) 0 1.5 - 1.5 Nitrate-N( mg/L) 10 10 10 50 Iron( mg/L) 0.3 0.3 0.3 0.3 - - - - 0.05 0.01 0.01 0.5 - 0 0 0 Nil Nil Nil Nil hardness 1000 250 mg/L(as CaCO3 ) Phosphate( mg/L) Arsenic( mg/L) Total coliform (MPN/100mL) E.coli (MPN/100mL) 6
  14. 14. 1.2. STATEMENT OF THE PROBLEM: Water quality has a direct impact on public health. More than 80% of deaths is caused due to water borne diseases. The water supply system in Kathmandu valley is insufficient as per demand of consumers due to centralization of Nepalese population day by day. The people of Kathmandu valley show an increasing trend of using jar water, mostly driven by the unreliable and quality compromised tap water supply and in part due to the perception and expectation of pure and safe drinking water. With the increasing demand and insufficient supply, it seems that in the near future, the urban dwellers would not have an option other than using bottled/jar water. Thus it’s high time to check the quality and monitor the bottled water industry. However, very few studies have been carried out to assess their quality and there are no agencies that regularly monitor their quality. 1.3. JUSTIFICATION: Safe drinking water is a fundamental right of human being. However, is the water that we drink safe? The answer is obviously “NO” as shown by the death statistics from water borne diseases which accounts to 80%. Driven by the perception of purity, people switch to buy bottled water. The question is not: why to check the quality of bottled water, it is: why not? People have the right to know the quality of water that they perceive to be pure. Hence, this case study is justifiable. 1.4. OBJECTIVES OF THE STUDY: 1.4.1. Broad Objective: To analyze the water quality of various brands of jar water marketed in the Kathmandu valley. 1.4.2. Specific Objectives: To analyze the Physio-chemical parameters of jar water sold in Kathmandu valley. To determine the bacteriological quality of jar water. 7
  15. 15. CHAPTER- 2 LITERATURE REVIEW Sharma S., (1978) analyzed the household drinking water in 39 localities of Kathmandu valley and found coliforms ranging from 4 to 460 cfu per 100 mL. DISVI (1989) conducted a study on the quality of drinking water of Kathmandu valley by taking 472 samples at 58 sampling points, 44 water taps, 7 storage, and 7 water treatment plants which showed existence of bacterial contamination in most of the sampling points. Ground water, a major source of drinking water in Kathmandu valley indicates high level of iron, magnesium and ammonia (JICA, 1990) ENPHO/DIVSI (1990) conducted a study on water quality of 21 stone spouts of Kathmandu city .The results showed heavy bacterial contamination in 81% of the total samples along with the presence of fecal contamination. ENPHO/DIVSI (1992) conducted a one year monitoring on microbiological quality of water supply in Kathmandu. Water samples were collected from 39 taps and 6 treatment plants.18% of the treatment plants and 50% of public taps showed significant contamination. Sharma S., (1993) studied the drinking water quality of Kathmandu and Pokhara. Significant contamination was observed. Coliform counts of 2400/100 mL and 4800/100 mL respectively in the sampled areas. The water supply system of Kathmandu is old and lack of maintenance has led to frequent malfunction (ADB, 1995). Masaaki N., and Hiroaki N., (1998) analyzed the bottled water in Kathmandu valley in July 1997. Bacterial contamination of the bottles purchased in Kathmandu (n=23) were checked using bacterial culture kit, "Test Paper for General Bacteria" and "Test Paper for Coliform" (SAN Chemical Co. Ltd.), following the direction of the manufacturer, and found that 7 bottles (30.4%) contained general bacteria, coliforms were detected in 4 8
  16. 16. bottles (17.4%) and 5 out of 23 bottles were found to have less content, i.e., water had leaked while transportation. The conclusion drawn from this study was that water bottles sold in the developing countries were contaminated with bacteria quite frequently. Joshi, et al (1999) conducted a study of bottled water in Kathmandu valley. Twenty different brands of mineral water were analyzed for hygienic quality and chemical constituents. In the microbiological analysis, coliforms were detected in 3 samples with fecal coliforms detected in 1 sample and Salmonella spp in 2 samples. The conclusion drawn was that the bottled water is contaminated frequently. Pokhrel S.R., (2000) analyzed 42 samples of bottled water from 7 companies for Physiochemical as well as microbiological parameters. He found that the physio-chemical parameters were under the acceptable limit whereas, bacterial count up to 162 was found in Total Plate Count. In addition to this, yeast as well as coliform was also detected. Bittner A., et al (2000) carried out a study on the quality of drinking water of Kathmandu. Samples were taken from various sources like well, stream and treatment plants, all of which showed contamination. Hence it was concluded that most drinking water supplies in Kathmandu are microbiologically contaminated. Prasai T., et al, 2002, analyzed a total of 132 water samples collected from various sources. Among the total samples, 49 were from tube wells, 57 from wells, 17 from taps and 9 from stone spouts. The analysis was carried out for various water quality parameters. The results showed that 82.6% of drinking water samples crossed WHO standards. During the study, 238 isolates of enteric bacteria were identified, of which 26.4% were Escherichia coli. Centre for Science and Environment, India(2003) analyzed 26 samples of 13 bottled water brands and raw water samples in Mumbai and found that every samples showed pesticides concentration between 0.0007 to 0.0042 mg/L. The maximum concentration was 40 times higher than European Economic Community (EEC) standards. Ribeiro A., et al (2006), analyzed water quality from various sources in Portugal. The objectives of this study were to analyze the seasonal fluctuations of fungal contamination, and to trace the origin of the contaminating fungal populations with molecular biology 9
  17. 17. techniques in a bottled water company. He analyzed water from water tank, water filter and bottled water twice monthly for fungal growth and found significant fungal contamination. The dominant fungal genera in order of highest numbers isolated were: Penicillium, Cladosporium and Trichoderma followed by Aspergillus, Paecilomyces and others. He also observed that fungal contamination increased during the warmer seasons, especially May and June. Warner N.R. et al., (2007) studied drinking water quality in Kathmandu valley. Water was sampled from over 100 sources including municipal taps, dug wells, shallow and deep aquifer tube wells and stone spouts. They found that the most problematic were total coliform and E. coli which was present in94% and 72% of all water samples respectively. Contamination by nitrate, ammonia and heavy metals was more limited. Gyawali R.,(2007) conducted a study on Microbial and chemical quality of water available in Kathmandu with 6 samples of tap and river from Sundarighat upstream and found that the physiochemical parameters were below WHO standards except chloride. Also, bacteriological contamination was 900 cfu/100 mL in average. Thakuri B.M., (2008) Conducted a study on the quality of bottled water in Kathmandu, taking 10 different brands of bottled water available in the valley and found that most of the Physio-chemical parameters were under the limit of WHO (1994). Microbial analysis showed that most brands had satisfactory quality though few numbers of coliforms were present. Pandey B., (2009) analyzed the drinking water quality of Central Development Region, Nepal. He analyzed a total of 243 samples: 130 from ground water source and 113 from springs. 20 of the ground water sample exceeded WHO standards. In addition to this, he concluded that most of the springs and ground water sources were heavily contaminated with fecal coliform bacteria. 10
  18. 18. CHAPTER- 3 METHODOLOGY 3.1. SAMPLE COLLECTION, TRANSPORTATION AND PROCESSING Samples of nine brands of jar water of 20 litres capacity were collected randomly from various restaurants as well as from jar water selling shops. For analysis of physicochemical parameters, water sample was collected in PVC sampling bottle. Some parameters such as temperature, pH, chloride, dissolved oxygen (DO), hardness, alkalinity and free carbon dioxide were determined in site while other parameters were determined in the laboratory of Central Department of Environment Science T.U. For determination of iron contents, about 1ml conc. HCl was kept in the samples bottles before the collection of water sample, in order to preserve the samples in reduced state. Samples for bacteriological analyses were collected in sterilized bottle, stored in ice cold box and transported to laboratory and were processed within 6 hours of collection. 3.2. SAMPLING FREQUENCY: Water quality was analyzed twice for each brand of bottle water during months of January, February and March when the difference in daily temperatures and change in season proceeds. The methodologies used to analyze various parameters are described below. 3.3. ANALYSIS OF WATER SAMPLES 3.3.1. Analysis of physicochemical parameters of water samples: "Standard Methods for the examination of water and wastewater", (APHA, 1998) was followed to analyze most of the physicochemical parameters of water. 3.3.2. Analysis of microbial variables of water sample: Microbial analysis was carried out following the "Standard Methods for the examination of water and wastewater", (APHA, 1998) and "Chemical and Biological methods for water pollution studies", 18th edition, R.K. Trivedi and PK. Goel 1984". 11
  19. 19. Table 3.3.1.: Methodology of Physico-chemical parameters and microbial analysis S. No. 1 2 3 4 5 6 7 8 9 10 11 12 13 Parameter Temperature pH Conductivity DO Iron Total Hardness Total Alkalinity Chloride Free CO2 Phosphate Nitrate Ammonia Total Coli form & Faecal coliform Equipments/Methods Mercury Thermometer pH meter Conductivity meter Winkler’s iodimetric titration Spectrophotometer EDTA method Titration method Titration method Titration method Spectrophotometer Spectrophotometer Spectrophotometer MPN method 1. Temperature For the determination of the temperature, water was collected in a beaker. Mercury filled Celsius thermometer was inserted into the beaker and reading was noted. 2. pH pH was measured by automatic digital pH meter. The pH meter was first calibrated with a standard buffer solution. The glass electrode was washed with distilled water. Then glass electrode was dipped in the beaker containing water sample until the reading stabilized at a certain point. Then pH reading was noted down. 3. Conductivity The instrument used was digital conductivity meter. The conductivity meter was first calibrated with standard Potassium chloride solution of 0.01N. Then reading was noted. 4. Chloride Chloride was measured by titration method. 50 mL of sample in a conical flask was taken. 2 mL of Potassium chromate was added to the sample solution. It was titrated against 0.02N silver nitrate until a persistent brick red color was appeared which was the end point of the titration. A blank by placing 50 mL of chloride free distilled sample water was also conducted. 12
  20. 20. Calculation Chloride (mg/L) = (a-b) × N ×35.5 × 1000 V Where, a = Volume of titrant (silver nitrate) for sample b= Volume of titrant (silver nitrate) for blank V = Volume of the sample in mL N = normality of silver nitrate 5. Total hardness Hardness is caused by the calcium and magnesium ions present in water. Total hardness was determined by EDTA method. This was done by titrating 100mL of sample in a conical flask and adding 1mL of buffer solution with Erichrome Black-T indicator against standard EDTA (Ethylene diamine tetra acetic acid). The solution was changed from wine blue at the end point. Total hardness might be caused by the sum of all metallic cations other than alkali metals and expressed as equivalent calcium carbonate concentration. Total hardness (as CaCO3), (mg/L) = mL of EDTA used×100 mL of sample 6. Calcium hardness Calcium hardness was determined by the same procedure as total hardness. Taking 50mL sample in a conical flask with 2mL of NaOH solution of 1N was titrated against EDTA solution using murexide indicator. At the end point, pink color changed to purple. Calcium, mg/L (as CaCO3) = Vol. of EDTA×N × 40.08 × 1000 Vol. of sample 7. Magnesium hardness Magnesium salts occur in significant concentration in natural waters which may be calculated as the difference between total hardness and calcium hardness. Magnesium hardness, mg/L (as CaCO3) = Total hardness – Calcium hardness 8. Free CO2 Free CO2 in water can be determined by using titration method. For this 100 mL of sample was taken in a conical flask and 2 drops of phenolphthalein indicator was added. Then it was titrated against 0.05N of NaOH from the burette until pink color was just appeared. 13
  21. 21. Calculation Free CO2 (mg/L) = A × Normality of NaOH × 44 ×1000 Volume of sample in mL Where, A = Volume of NaOH used in mL 9. Dissolved Oxygen ( Winkler’s Method ) DO was measured by using APHA, (1998) method. The sample was collected in a 300 mL BOD bottle carefully, avoiding any kinds of bubbling and trapping of air bubbles in the bottle after placing the stopper. To the 50 mL sample taken in the conical flask 2 mL of manganese sulphate (MnSO4) and 2 mL of Sodium azide solution was added well below the surface from the wall of the bottle. A precipitate was appeared. Then the stopper was placed tightly and the bottle was shaken by inverting the bottle repeatedly to insure proper mixing of the contents. The bottle was kept for some time to settle down the precipitate, 2 mL of conc.H2SO4 was added to it and shaken well to dissolve all the precipitate. Then 50 mL of sample were taken in a conical flask and titrate against sodium thiosulphate (Na2S2O3) of 0.025N using starch as an indicator. At the end point the initial blue color changed to colorless. Calculation: DO (mg/L) = (mL× N) of titrant ×8 ×1000 V2× (V1-V) V1 Where, V1= volume of sample bottle after placing the stopper V2= volume of part of content titrated V= volume of MnSO4 and KI added 10. Total Alkalinity Total Alkalinity is the measure of the capacity of the water to neutralize a strong acid. The alkalinity in water is generally imparted by the salts of carbonates, bicarbonates, phosphates, nitrates, borates, silicates etc. together with hydroxyl ions in free state. Total alkalinity of water was determined by titrimetric method. 100mL sample in a conical flask with 2-3 drops of methyl orange was titrated against standard, 0.02N H2SO4. At the end point, yellow color was changed to pink color. 14
  22. 22. Total Alkalinity (mg/L) = a ×N×1000×50 mL of sample where, a= Volume of standard H2SO4 consumed in titration N= Normality of H2SO4 used 11. Phosphate – P Phosphate content in the given water sample was determined as inorganic phosphate by calorimetric method. In this method, 50mL of the filtrate clear sample was taken in a conical flask. 20 mL of ammonium molybdate was added to it. 5 drops of SnCl 2 solution was added to it. The solution becomes blue and the reading was taken at 690 nm on the spectrometer within 10-12 minutes. Same procedure was repeated for the standard solution of different concentration for distilled water. The concentration was determined with the help of standard curve obtained by plotting standard values against absorbance. 12. Nitrate-N Nitrate content in the water sample was determined by Phenol disulphonic acid method. In this method, 50 mL of filtrate sample was taken in a porcelain basin and was evaporated to dryness. It was cooled and residue was dissolved in 2 mL of phenol disulphonic acid and was diluted to 50 mL. 6 mL of liquor ammonia was added to develop yellow color. Then the reading was taken at 410 nm on spectrophotometer. Same procedure was repeated for the standard solution of different concentration and for distilled water. Then the concentration of Nitrate-N was determined from the standard curve obtained by plotting standard value against absorbance. 13. Ammonia-N Ammonia content in a water sample was determined by colorimetric method. In this method 100 mL of water sample was taken in a volumetric flask. 1 mL of ZnSO4.7H2O was added, 1 mL of 10% NaOH was added into it. Then it was stirred and filtered. One drop of 50% EDTA was added and well mixed. Then 2 mL of Nessler’s reagent (K2HgI4) was added. Then the reading was taken at 420 nm on spectrophotometer. The same procedure repeated for the standard solution of different concentration. Then the concentration determined with the help of standard curve. 15
  23. 23. 14. Total Iron Iron content in the water sample was determined by colorimetric method. In this method 50 mL of the sample was taken in a conical flask. 2 mL of concentrated HCl and 1 mL of hydroxylamine hydrochloride solution was added. Then some glass beads were put in the flask and boil till the content is reduced about half. It was cooled and 10 mL of acetate buffer solution and 2 mL of Phenothroline solution was added, then orange red color was appeared. Distilled water was added to make the volume 100 mL in a volumetric flask. Let it stand for 10 minutes, and the absorbance of the color was measured by using spectrophotometer at 510 nm using distilled water blank with the same amount of chemical. The same procedure was repeated for standard solution of different concentrations. Then the concentration was determined with the help of standard curve. 15. MPN Test: The purity of drinking water regarding bacterial contamination is evaluated by testing the presence of coliforms as these are designated as indicator organism of faecal contaminations. Coliform bacteria in the water sample were determined by Most Probable Number (MPN) or Multiple - Tube fermentation test. This test is performed sequentially in 3 stages: 1) Presumptive coliform test: This is used to detect coliforms in a water sample. In this test lactose fermentation tubes (Mac-Conkey Broth) were inoculated with different water volumes and production of acid and gas from the fermentation of lactose within 48 hrs in any of the tubes was the presumptive evidence of coliforms in the water sample. a) 10 mL of water sample was inoculated in each of 3 tubes containing 10 mL of the double strength of Mac-Conkey broth. b) 1 mL of water sample was inoculated in each of 3 tubes containing 9 mL of the single strength of Mac-Conkey broth. c) 0.1 mL of water sample was inoculated in each of 3 tubes containing 9.9 mL of the single strength of Mac-Conkey broth. All the inoculated tubes were incubated at 37° C for 24 hours. After incubation, the tubes were counted which had produced both acid and gas. The tubes showing 16
  24. 24. negative result were further incubated at 37° C for 24 hours. The tubes showing gas formation and acid formation were further inoculated for confirmatory test. 2) Confirmed coliforms test: This test is used to confirm the presence of coliforms and to determine the MPN value in water sample. In this test water samples from all the positive presumptive Mac Conkey broth tubes were inoculated into two sets of tubes of BGLB (Brilliant Green Lactose Bile salt) broth and one set incubated at 37oC for 24-48 hours- for total coliform and another incubated at 440 C in water bath for 24 hours for fecal coliform. Positive confirmed tubes were used to determine MPN/100ml by following statistical Method (MPN table). 3) Completed coliforms test: This test is used to establish the presence of Total Coliform as MPN/100 ml in a water sample. Positive tube from the confirmatory was streaked on the plate of EMB Agar and incubated it at 37° C for 24 hours. After 24 hours, the colonies that had typical growth (dark centre with greenish metallic sheen) and atypical growth were transferred to nutrient agar slant and Mac Conkey Broth and incubated at 37° C for 24 hours. The presence of total coliform and faecal coliform bacteria was confirmed by Gram Staining and Spore Staining Technique. 17
  25. 25. CHAPTER- 4 RESULTS AND DISCUSSION The table with respective values of each of the parameters is shown in the annex. 1. pH: Fig 4.1: Comparison between pH values pH is the negative logarithm of hydrogen ion concentration. It is used to express the intensity of acidic or alkaline condition of a solution. The pH values in all the samples tested during Jan / Feb. and Feb. / March range from 6.5-7. pH of pure water is 7. pH is an extremely important variable because it is the controlling factor determining the solubility of most metals and also because most micro-organisms can survive within a narrow range of pH. pH is also an important factor in water treatment. Proper chemical treatment of water including disinfection requires pH control. The values of pH obtained are within the WHO standards of 6.5-8.5. 18
  26. 26. 2. Temperature: Fig 4.2: Graph showing temperature values Temperature of fresh water varies normally from 0 to 35ºC depending on the source, depth and season. The temperature of water affects some important physical properties and characteristics of water such as density, viscosity, conductance, salinity, solubility of dissolved gases etc. Also, chemical and biological reaction rates increase with temperature. In the above graph, the temperature in the first test during January/February shows range between 15-18 ºC and in the second test conducted during February/March, the ranges are between 19-23 ºC. 3. Conductance: Fig 4.3: Comparison of conductance values in two tests 19
  27. 27. Electrical conductivity is the measure of the capacity of water to conduct electric current. The values of conductance range from a minimum of 15µs/cm for brand 3 to a maximum of 815µs/cm for brand 4 in first test. Similarly in the second test minimum value is 70 µs/cm for brand 3 and maximum value is 768 µs/cm for brand 4. In comparison of other brands, brand 4 has high conductance. High values of conductance indicate high dissolved gases and other chemicals in the water. There is no guideline value for conductivity; however, values above 400us/cm may affect the chemical quality of drinking water. 4. Alkalinity: Fig 4.4: Comparison of alkalinity values in two tests Alkalinity is also a major parameter affecting water quality that mainly acts for pH neutralization. Alkalinity measurements are used as the means for evaluating the buffering capacity of water. The minimum values of alkalinity were 15mg/L for brand 2 and 25 mg/L for brand 3 in the 1st and 2nd tests respectively, while the maximum values were 90 mg/L for brand 7 and 142 mg/L for brand 5 in the 1st and 2nd tests respectively. These values are well below the permissible limits. However, there is an increasing trend of alkalinity values. In natural water, most of the alkalinity is caused by CO2. Since the concentration of free CO2 also has increased in the second test, the rise in the values of alkalinity in the 2nd test seems to be obvious. 20
  28. 28. 5. Hardness: Fig 4.5: Comparison between the hardness values Hardness is imparted to the water mainly by calcium and magnesium ions. Calcium is essential element for human beings (nearly 2 gm per day) and plant growth. However, hard water is generally undesirable because it forms precipitate with soap, produces scales in boilers on heating and has high boiling point due to which it is unsuitable for cooking. The minimum values of total hardness were 10mg/L for brand 8 and 8 mg/L for brand 2 and 8 in the 1st and 2nd tests respectively, while the maximum values were 80 mg/L for brand 7 and 76 mg/L for brand 5 in the 1st and 2nd tests respectively. The WHO standard for hardness is 200mg/L. Thus all the values are within acceptable limits. The minimum values of Calcium hardness were 6mg/L for brand 8 and 4 mg/L for brand 2 in the 1st and 2nd tests respectively, while the maximum values were 46mg/L for brand 7 and 42 mg/L for brand 5 in the 1st and 2nd tests respectively. Similarly, for magnesium hardness, the maximum values were 34 mg/L for brand 7 and 34 mg/L for brand 5 in the 1st and 2nd tests respectively and minimum values for 1st and 2nd test were 4 mg/L for brand 8 and 2 mg/L for brand 8 respectively. 21
  29. 29. 6. Free CO2: Fig 4.6: Graph showing values of Free CO2 in two tests Surface water normally contains less than 10mg/L of free CO2 while some ground water may contain 30-50mg/L of free CO2. High concentration of free CO2 indicates pollution from domestic sewages and industries. However, there are no prescribed limits of free CO 2 for drinking water as free CO2 do not bring about physiological effects. The values for free CO2 for most of the samples are zero in the 1st test except brands 6, 7, and 8 whereas in the 2nd test. All the sample show significant values with a maximum of 48.4 mg/L for brand 8 and a minimum of 6.6 mg/L for brand 5. 7. Chloride: Fig 4.7: Graph showing chloride values in two tests 22
  30. 30. Chloride is present in appreciable amounts in all natural water. Concentration varies from few milligrams to several thousand milligrams per liter. High concentration of Chloride may indicate pollution of organic origin, as well as results in corrosivity and impaired taste. The permissible limit of chloride according to WHO is 250 mg/L. Drinking water is often chlorinated for disinfection. The concentration of chloride is high for brand 4 in both the months. The values of chloride range from 1.42-86.62 mg/L in the 1st test and 8.5293.72 mg/L in the 2nd test. All the values are within WHO guidelines. 8. Dissolved Oxygen (DO): Fig 4.8: Comparison between DO values Oxygen is dissolved in water in varying concentrations. It is a very important water quality parameter and is also an index of physical and biological processes going on in water. Analysis of DO is very important in water pollution control. The guideline value for DO is >5 mg/L according to WHO. Brand 9 shows higher DO values among all. The obtained values are in the range 7.7-8.2 mg/L in the 1st test and 6.9-9.3 mg/L in the 2nd test which satisfy WHO standards. 23
  31. 31. 9. Total Iron: Fig 4.9: Graph showing the values of Total Iron Iron has got a little concern as health hazard, but it still is considered as a nuisance in excessive quantities. High iron content produces bitter and astringent taste. The WHO has set the permissible limit of iron to 0.3 mg/L. Among all brands, brand 9 has the highest iron concentration in both the months. The values of iron range from 0.014-0.1 mg/L in the first test to 0.015-0.12 mg/L in the 2nd test which all lies below the acceptable limits. 10. Nitrate- N: Comparision between nitrate values 0.025 0.02 mg/L 0.015 0.01 Nitrate A(mg/L) 0.005 Nitrate B(mg/L) 0 1 2 3 4 5 6 7 8 9 Brands Fig 4.10: Comparison between Nitrate-N values Nitrates are present in trace amounts in surface water but in some ground water, nitrates may be high. High nitrite and nitrate concentration in water causes a disease called methemoglobinaemia. In drinking water, nitrate concentration should be less than 10 mg/L 24
  32. 32. according to WHO. The values obtained are in the range of 0.01-0.02 mg/L in both tests, all below the permissible limits. 11. Phosphate-P: Fig 4.11: Graph showing values of Phosphate-P Though in low concentration, phosphate is an important nutrient present in water. The values of phosphate range from 0.14-0.32 mg/L in the first test and 0.15-0.29mg/L in the 2nd test. The highest value of phosphate is seen for brand 9 in both the months. 12. Ammonia-N: Comparision between ammonia values 0.012 0.01 Ammonia A(mg/L) 0.008 mg/L Ammonia B(mg/L) 0.006 0.004 0.002 0 1 2 3 4 5 6 7 8 9 Brands Fig 4.12: Comparison between ammonia-N values Ammonia is generally an indication of pollution in drinking water. According to the guideline value given by WHO, the concentration of ammonia should be 0 mg/L. The maximum concentration of ammonia was found to be 0.01 mg/L and in most of the test was undetectable, that means, less than 0 mg/L. 25
  33. 33. 13. Bacteriological examination: Table 4.13.1 Bacteriological examination water during January/February Brands Presumptive count/100 mL Total Coliforms 1 210 2 >1100 3 210 4 Nil 5 Nil 6 21 7 210 8 210 9 Nil In the presumptive count, out of nine brands, 6 brands were found to be contaminated with coliform. Brands 4, 5, and 9 were found to have no contamination. Table 4.13.2: Bacteriological examination water during February/March Brands Total coliform Fecal coliform (MPN/100mL) (MPN/100mL) 1 >1100 4 2 >1100 4 3 240 4 4 9 Nil 5 210 3 6 15 Nil 7 >1100 20 8 >1100 7 9 Nil Nil 26
  34. 34. In this test, only one brand was found to be devoid of both total and fecal coliform while two other brands showed total coliform but no fecal coliforms and the rest brands showed both total and fecal coliform. Among the samples, brands 1, 2, 7 and 8 were found to be heavily contaminated. Data Analysis: Analysis of physicochemical parameters during January/February showed that the pH of the samples range from 6.55 to 7.63. Temperature of the samples range from 16-18°C. Conductance was found to be as low as 15µs/cm for brand 3 and as high as 815 µs/cm for brand 4. The value of alkalinity was also found to be variable. It ranged from a minimum of 10 mg/L for brand 2 to 90 mg/L for brand 7. The values of hardness range from 10 to 68( as CaCO3 ).The values of Free CO2 was found to be 0 in most of the brands while brand 7 showed a maximum value of 17.6 mg/L. The values of chloride were also highly variable. The value was below 10 mg/L for 5 brands and a maximum of 86.62 mg/L was obtained for brand 4. The values of DO were more or less similar and ranged from 7.7 8.2 mg/L. The value of iron was obtained in a range of0.04-0.05 mg/L except brand 9 which showed a value of 0.10 mg/L. The values of nitrate obtained were in the range of 0.01 - 0.02 mg/L. Similarly the values of phosphate were in the range of 0.14 - 0.32 mg/L. Ammonia was not detected in any of the samples. During Jan/Feb, only three samples of brands 4, 6, and 9 have no coliforms at all. That means 66% of the sample is heavily contaminated. According to the WHO limit for the Presumptive count, greater than 10 presumptive count/100mL are unsatisfactory. Due to unforeseen circumstances as well as physical constraints, completion of MPN test was hindered. Similarly, Analysis of physicochemical parameters during February/March showed that the pH of the samples range from 6.81 to 7.89. Temperature of the samples range from 19 -23°C. Conductance was found to be as low as 70µs/cm for brand 3 and as high as 768 µs/cm for brand 4. The value of alkalinity was also found to be variable. It ranged from a minimum of 25 mg/L for brand 3 to 142 mg/L for brand 5. The values of hardness range from 8 to 76(as CaCO3).The values of Free CO2 was found in a range between 6.6 - 48.4 mg/L. The values of chloride were 8.52 mg/L for brand 1 and a maximum of 93.72 mg/L 27
  35. 35. was obtained for brand 4. The values of DO ranged from 6.9 – 9.3 mg/L. The value of iron was obtained in a range of 0.05-0.07 mg/L except brand 9 which showed a value of 0.12 mg/L. The values of nitrate obtained were in the range of 0.01 - 0.02 mg/L. Similarly the values of phosphate were in the range of 0.15 - 0.29 mg/L. Ammonia was not detected in any of the samples. During Feb/Mar, with the onset of summer, the MPN count/100 mL has also increased. Total coliform was found to be Nil in only one sample (Brand 9) whereas fecal coliform were found to be nil in 3 samples, Brand 4, 6 and 9. In the second test, total coliforms were present in 89% of the total sample and fecal coliform was present in 66% of the total samples. Brand 4 and 5 showed no contamination in the first test but showed contamination in the second test. This may be partly because of increased microbial activities with the onset of summer or due to the contamination of jar. Even two jars of same brand may vary in quality since during refilling and processing, contamination may occur. Similarly, there is no assurance whether these jars are even processed. The results show that the samples are heavily contaminated with coliforms. It may be due to the improper water processing techniques as well as jars. Furthermore, the source of jar water is not mentioned in any of the jars. 28
  36. 36. CHAPTER- 5 CONCLUSION AND RECOMMENDATION 5.1. CONCLUSION: Hence from the above results, it can be concluded that jar water, although thought to be pure, cannot be relied upon for its safety. Various physio-chemical parameters like DO, hardness, total iron, phosphate, nitrate, ammonia, alkalinity, pH, etc were analyzed using standard methods of APHA (1998). All the Physio-chemical parameters like DO, hardness, alkalinity, pH were within WHO acceptable limits. Ammonia was detected in some samples but is within the limits set by WHO. Iron and Nitrates were found in small quantities but within the limits set by WHO. With the onset of summer, the concentration of some parameters rose like Free CO2, conductance, alkalinity etc, which are more or less affected by temperature. Thus from the physio-chemical aspect, the quality of water is good. From the microbiological point of view, 66% of the total samples were heavily contaminated with total coliforms in the first test during January/February. Three brands of jar water tested had no coliforms at all. During the second test in February/March, 89% of the total sample was found to be contaminated with total coliforms whereas 66% were contaminated with fecal coliforms. Total coliforms as many as 1100 MPN/100mL and a maximum of 20 MPN/100 mL of fecal coliforms were enumerated. Hence, it can be concluded that the water samples are heavily contaminated with coliform bacteria and unsatisfactory for drinking purpose. As per the NRDC (1999) result says, “While much tap water is indeed risky, having compared the available data, we conclude that there is no assurance that bottled water is any safer.” Similar is the conclusion of this study, that there is no assurance that since water comes out of a bottle does not mean it is free from contamination. 29
  37. 37. 5.2. RECOMMENDATIONS: 1. There are varieties of jar water and their quality also varies. Thus it is necessary to pick up the right brand. 2. Stricter rules should be made and implemented to regularly monitor the bottled water qualities. 3. The labels of bottled water must include not only the pristine glaciers and Himalayan springs but also the relative concentrations of water quality parameters. 4. Since jar water are reused, sometimes they are used up to the extent that there is neither company name nor any labels. In such condition, the jar may itself contaminate the water although the water is safe. 5. All the bottled water companies should fulfill the basic water quality standards given by the Government of Nepal and then registered to NS Standards since only three companies have done it so far. 6. Awareness should be created to public for either using disinfectants or boiling water before use rather than rely on the belief of purity. 30
  38. 38. REFERENCES ADB(2004) Water for all: The impact of water on the poor, Asian Development Bank, Manila. APHA. ,1998: Standard methods for the examination of water and waste water. 20th edition, American Public Health Association, Washington D.C. 1-47 pp. Bittner A., Halsey T., Khayyat A., Luu K., Maag B., Sagara J., and Wolfe A., (2000), “Drinking Water Quality and Point – of – use Treatment Studies in Nepal” CBS. 2001. Statistical Year Book of Nepal HMG, Central Bureau of Statistics. Chapagain, A.K., and Hoekstra, A.Y. (2004) Water Footprints of Nation: Volume I: Main Report, UNESCO- IHE, Institute for Water Education, Value of water: Research Report Series no. 16, pp. 75. Diwakar J. (2007), “Assessment of Drinking Water Quality of Bhaktapur Municipality, M.Sc. Thesis, Central Department of Environment Science, TU. ENPHO (2001), “Drinking Water Quality and Sanitation Situation in UNICEF’s project area: Kavre, Parsa and Chitwan, September 2001.” Government of Nepal, National Drinking Water Standards, 2062 and National Drinking Water Quality standard Implementation Guideline, 2062 year: 2063 B.S. Government of Nepal, Ministry of Physical Planning and Works, Singhadurbar, Kathmandu, Nepal. Gyawali R.,(2007), “ A Study on Microbiological and Chemical Quality of Water of Kathmandu” M.Sc. Thesis, Central Department of Microbiology, TU. Masaaki N., and Hiroaki N., (1998), “Quality of the bottled water in Nepal”, Japanese Journal of Mountain Medicine, Vol.18, pp 107-110. 31
  39. 39. Pandey B., (2009), “A Case Study of Drinking Water Quality Status in Central Development Region, Nepal” Pimentel D., Berger B., Filiberto D., Newton M., Wolfe B., Karabinakis E., Clark S., Poon E., Abbett E., and Nandagopal S.,(2004) Water Resources, Agriculture and the Environment, Report 04-1, Cornell University, College of Agriculture and Life Sciences Prasai T., Lekhak B., Joshi D.R.,and Baral M.P., “Microbiological Analysis of Drinking Water of Kathmandu Valley”, Nepal Academy of Science and Technology, Kathmandu, Nepal. Ribeiro, A., Machado, A.P., Kozakiewicz, Z., Ryan M.,Luke B., Buddie A.G., Venancio A., Lima N.and Kelley J. (2006), “Fungi in Bottled Water, ACase Study of a Production Plant.” Shrestha RR, Sharma S., Bacteriological Quality of Drinking Water in Kathmandu City: A Review of ENPHO/DISVI Reports, 1988-1992, (Katmandu: Environment and Public Health Organization, 1995) Thakuri B.M., (2008), “Analysis of Bottled Water Available in Kathmandu Valley”,M.Sc. Thesis, Central Department of Environment Science, TU. The World’s Water, “The Biennial Report on Fresh Water Resources: 2004-2005”, Island Press United Nations (2003), “Water for People, Water for Life: A Joint Report by the 23 UN Agencies concerned with Fresh Water, The UN World Water Development Report. WHO. 1993. Guidelines for Drinking Water Quality, Volume I, II and III, World Health Organizations, Geneva. WHO. 1996. Guidelines for Drinking Water Quality 2nd edition. Volume II, Health criteria and other supporting information. World Health Organizations, Geneva. 32
  40. 40. ANNEX Results of the Physio-chemical tests: Table- Physio-chemical parameters of jar water during January/February Sample(A) Brand Brand Brand Brand Brand 5 Brand Brand Brand 7 8 9 6 Brand 1 2 3 4 pH 6.74 7.15 6.62 7.63 7.26 6.55 7.38 6.89 7.41 Temperature(º 16 17 16 17 15 17 17 16 18 Conductance 150 20 15 815 96 85 223 168 50 Alkalinity 45 10 15 55 65 40 90 45 15 (as 36 18 36 68 18 34 80 10 16 Ca-hardness (as 30 10 18 42 12 22 46 6 10 8 18 26 6 12 34 4 6 Parameters C) (mg/L) Hardness CaCO3) CaCO3) Mg-hardness (as 6 CaCO3) Free CO2 (mg/L) 0 0 0 0 0 4.4 17.6 4.4 0 Chloride(mg/L) 17.04 1.42 2.84 86.62 5.68 8.52 15.9 14.2 9.94 DO(mg/L) 7.7 7.7 7.7 8.2 8.1 8.1 8.1 7.7 8.1 Iron (mg/L) 0.05 0.04 0.05 0.05 0.04 0.05 0.04 0.05 0.10 Nitrate A(mg/L) 0.01 0.01 0.01 0.02 0.01 0.01 0.02 0.01 0.02 Phosphate(mg/L) 0.17 0.15 0.14 0.16 0.18 0.14 0.23 0.18 0.32 Ammonia 0.01 0.01 ND 0.01 0.01 ND 0.01 ND 0.01 A(mg/L) ND=Not Detected 33
  41. 41. Table- Physio-chemical parameters of jar water during February/March Sample(B) Brand Brand Brand Brand Brand 5 Brand Brand Brand 7 8 9 6 Brand 1 2 3 4 pH 7.12 7.35 7.43 7.89 7.85 6.81 7.38 7.66 7.83 Temperature(º 20 22 20 23 22 19 21 21 21 Conductance 187 195 70 768 118 106 295 253 122 Alkalinity 55 35 25 125 142 60 130 70 40 (as 40 8 44 54 76 44 12 8 24 Ca-hardness (as 22 4 36 32 42 28 8 6 14 4 8 22 34 16 4 2 10 Parameters C) (mg/L) Hardness CaCO3) CaCO3) Mg-hardness (as 18 CaCO3) Free CO2 (mg/L) 22 11 13.20 41.8 6.6 28.6 19.8 48.4 24.2 Chloride(mg/L) 8.52 21.3 28.82 93.72 9.94 11.36 11.36 21.3 25.56 DO(mg/L) 8.1 7.3 7.7 6.9 7.3 6.9 8.1 7.3 9.3 Iron (mg/L) 0.05 0.05 0.06 0.05 0.06 0.05 0.05 0.07 0.12 Nitrate B(mg/L) 0.01 0.01 0.02 0.02 0.01 0.01 0.02 0.01 0.02 0.15 0.17 0.16 0.19 0.15 0.15 0.17 0.21 0.29 0.01 0.01 0.01 ND 0.01 0.01 ND 0.01 ND Phosphate(mg/L) Ammonia B(mg/L) ND=Not Detected 34
  42. 42. MPN DETERMINATION FROM MULTIPLE TUBE TEST 95 PERCENT NUMBER OF TUBES GIVING MFN CONFIDENCE LIMITS POSITIVE REACTION OUT OF Index per 100 3 of 10 3 of 1 3 of 0.1 Lower Upper ml. ml. ml. ml. each each each 0 1 <0.5 9 0 3 1 0 <0.5 13 0 3 0 0 <0.5 20 1 4 0 1 1 21 1 7 1 0 1 23 1 7 1 1 3 36 1 11 2 0 3 36 1 11 0 0 1 36 2 9 0 1 3 37 2 14 1 0 3 44 2 15 1 1 7 89 2 20 2 0 4 47 2 21 2 1 10 150 2 28 0 0 4 120 3 23 0 1 7 130 3 39 0 2 15 380 3 64 1 0 7 210 3 43 1 1 14 230 3 75 1 2 30 380 3 120 2 0 15 380 3 93 2 1 30 440 3 150 2 2 35 470 3 210 3 0 36 1,300 3 240 3 1 71 2,400 3 460 3 2 150 4,800 3 1,100 From: Standard Methods for the Examination of Water and Wastewater, Twelfth edition. (New York: The American Public Health Association, Inc., p. 608.) WHO Standards For Presumptive Coliform Count: Class I II III IV Grade Excellent Satisfactory Suspicious Unsatisfactory Presumptive count/100mL 0 1-3 4-10 >10 35 E. coli count/100mL 0 0 0 0, 1, or more
  43. 43. Composition of Eosin Methylene Blue (EMB) Peptic digest of animal tissue 10.00 gm / Ll Dipotassium phosphate 2.00 gm / L Lactose 5.00 gm / L Sucrose 5.00 gm / L Eosin – Y 0.40 gm /Ls Methylene Blue 0.65 gm / L Agar 13.50 gm / L Final pH at 25ºC → 7.2 ± 0.2 Composition of Nutrient Agar Peptic digest of animal tissue Beef extract Yeast extracts Sodium Chloride Agar Final pH at 25ºC → 7.2 ± 0.2 5 gm / L 1.5 gm / L 1.5 gm / L 5.0 gm / L 15.0 gm / L Composition of Mac Conkey Broth Peptic digest of animal tissue 20 gm / L Lactose 10 gm / L Bile salt 5 gm / L Sodium Chloride 5 gm / L Neutral Red 0.075 gm / L Final pH at 25ºC → 7.2 ± 0.2 Composition of Brilliantly Green Lactose Bile Growth (BGLB) Peptic digest of animal tissue 10 gm / L Lactose 10 gm / L Oxgall 20 gm / L Brilliant Green 0.0133 gm / L Final pH at 25ºC → 7.2 ± 0.2 36

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