Environ Monit Assess (2011) 178:477–485
DOI 10.1007/s10661-010-1706-y
Ground water quality in the Kathmandu valley of Nepal
Bhoj Raj Pant
Received: 2 November 2009 / Accepted: 6 September 2010 / Published online: 21 September 2010
© Springer Science+Business Media B.V. 2010
Abstract A study was undertaken to assess the
quality of groundwaters in the Kathmandu Valley,
Nepal. The groundwater samples were randomly
collected from shallow well, tube well, and deep-
tube wells located at different places of Kathmandu,
Lalitpur, and Bhaktapur districts in the Kathmandu
valley. Physical, chemical, and microbiological pa-
rameters of the samples were evaluated to estimate
the groundwater quality for drinking water. It was
found that the groundwater in the valley is vulner-
able to drink due to presence of iron and coliform
bacteria. Iron was estimated to be much higher
then the acceptable limit of World Health Organi-
zation (WHO) drinking-water quality guidelines
(1.9 mg/L). Total coliform bacteria enumerated in
groundwaters significantly exceeded the drinking-
water quality standard and observed maximum
coliform (267 CFU/100 mL) in shallow wells. The
electrical conductivity and turbidity were found to
be 875 μS/cm and 55 NTU, respectively, which are
above the WHO recommendations for drinking
water guidelines. However, pH value was mea-
sured within the acceptable limit. Arsenic, chlo-
ride, fluoride, and hardness concentrations were
B. R. Pant (B)
Nepal Academy of Science and Technology,
G.P.O. Box 3323 Kathmandu, Nepal
e-mail: bhojrajpant@yahoo.com
found to be in agreement with the recommenda-
tions of WHO drinking-water quality guidelines.
Keywords Kathmandu valley · Groundwaters ·
Sampling · Coliform · Water quality
Introduction
Kathmandu valley has strongly increasing water
demand due to rapid growth of urban population.
Treated water supply is limited and insufficient
to meet increased water demand for domestic
uses. In this circumstance, groundwaters may be
a potential source for drinking water in the valley.
In Kathmandu valley, about 45% people living in
urban or semi-urban areas depend on groundwa-
ters for drinking and other domestic requirements
(MOPPW 2003). Generally speaking, groundwa-
ter is characterized by low temperature, low re-
dox potential, high carbon dioxide and mineral
content, less amount of suspended solids, and free
from microbial contaminants. By satisfying the
optimum drinking water requirements, groundwa-
ter is often considered as the preferred source for
public water supply and is also widely explored for
private, domestic, and industrial uses.
Recently, rapid urbanization and industrial-
ization have induced profound effects on urban
groundwater quality in Kathmandu valley, which
478
are inextricably linked with water supply pro-
vision and effluent disposal practices. Ground-
water pollution in urban areas is mostly due
to infiltration of urban storm water, leakage of
wastewaters and septic reservoirs, and improper
industrial activities (Robertson et al. 1991; Jeong
2001; Rivett et al. 2002; Dechesne et al. 2004). The
wastewaters and septic system effluent contain
high concentration of dissolved organic carbon,
ammonia, pathogens, and organic micro pollu-
tants as well as heavy metals and trace elements.
Percolation of such pollutants to groundwater ta-
ble greatly influences the water quality in ground-
water aquifer.
In this work, the groundwater quality of
Kathmandu valley was evaluated for drinking pur-
pose and estimated the pollutants level in the
groundwater aquifer. Physical parameters such as
temperature, pH, and electrical conductivity were
measured in situ. Analysis leading to the esti-
mation of turbidity, hardness, chloride, alkalinity,
iron, arsenic, and fluoride was performed in the
laboratory. Microbiological analysis for total col-
iform count was also carried out.
Fig. 1 Map of
Kathmandu valley
showing sampling sites in
Kathmandu, Lalitpur and
Bhaktapur
Environ Monit Assess (2011) 178:477–485
Study area
The study area covers urban and semi-urban regions
in the Kathmandu valley (Fig. 1). The Kathmandu
valley contains three major cities: Kathmandu,
Lalitpur, and Bhaktapur with approximate pop-
ulation of 1.6 million (CBS 2001). The valley is
located in the midland of the Himalayas, lies in
between 27◦ 32 and 27◦ 49 North and 85◦ 12 and
85◦ 32 East is almost round in shape with a diam-
eter of approximately 30 km E–W and 25 km N–
S (Dill et al. 2001). The valley has a central flat
part at an elevation of 1,300–1,400 m above mean
sea level and covers an area ∼900 km2 (Sharma
1997) with an average population density of 1,800
persons per km2 (CBS 2001).
It is difficult to enumerate exact number of
people who have been using groundwaters from
these sources. Nevertheless, the interview con-
ducted during sampling estimates, about 0.5 mil-
lion people directly or indirectly, depends upon
these groundwaters to satisfy their daily water
demand. The groundwater in the valley at present
is under threat because of heavy water withdrawal
Environ Monit Assess (2011) 178:477–485
and increased water pollution level. For the past
few years, groundwater in the valley is being
heavily used for household purpose and also for
other applications which resulted in decline of
the groundwater level (SOE 2001). According to
Stanley and Macdonald (1994), sustainable limit
of groundwater withdrawal is approximately 26
million liters per day and should not exceed 40
million liters (NWSC 1992). Nevertheless, total
groundwater extracted per day in the Kathmandu
valley at present is approximately 59 million liters
(Metcalf and Eddy 2000), which is much higher
than the permitted limit. The valley is in a warm
temperate climate zone and receives an annual
average rainfall of approximately 1,900 mm with
temperatures ranging from approximately 0◦ C in
the winter up to 30◦ C in the summer (NWSC
1992).
Materials and method
Hydrochloric acid, sulfuric acid, hydroxylamine
hydrochloride, ammonium acetate, acetic acid, fer-
rous ammonium sulfate, phenolphthalein, EDTA,
phenonthroline (Qualigens fine chemicals), ammo-
nium chloride, potassium permanganate, potassium
chromate, silver nitrate, methyl orange (Merck),
and magnesium sulfate (Ranbaxy) were the AR
grade chemicals purchased from local suppliers in
Kathmandu. Membrane filter (Whatman) and M-
endo agar Hi-media were used in the estimation
of total coliform. Polyethylene bottles (∼250 mL)
were used for sampling. Standard arsenic test
kits (Macherey–Nagel) were purchased from local
enterprises.
Sampling
The groundwater samples were randomly col-
lected from shallow wells of depth 3–11 m, tube
wells of depth 11–30 m, and deep-tube wells of
depth >30 m at different places of Kathmandu,
Lalitpur and Bhaktapur in between October and
December of 2004. Sampling was carried out accor-
ding to standard method described in the litera-
ture (Greenberg et al. 1992). Polyethylene bottles,
thoroughly cleaned by distilled water, were used
for sample collection. Samples to be analyzed for
479
total coliform bacteria were collected in the bot-
tles sterilized in an autoclave at 121◦ C and 15 LB
pressure for 15 min. Samples for the analysis of
chemical parameters (hardness, chloride, alkalin-
ity, iron, arsenic, and fluoride) were collected in
polyethylene bottles cleaned by distilled water for
several times. Before collecting the samples, the
sample bottles were purged at least three times by
the water to be collected from respective ground-
water sources. Dip sampler was used to collect
the samples from shallow wells while the samples
from tube well and deep-tube wells were collected
either by pumping through hand pump or by us-
ing electric motor. The samples were stored in a
portable icebox and transported to the laboratory
within 6 h and stored at ∼4◦ C in a refrigerator un-
til physical, chemical, and microbiological analysis
were carried out.
Sample analysis
The samples were analyzed for temperature, pH,
electrical conductivity (EC), turbidity, hardness,
chloride, alkalinity, iron, arsenic, fluoride, and to-
tal coliform bacteria. The temperature, pH, and
EC were measured in situ at sample collection
sites by using a portable thermometer (LINE
SEIKI TC-1100), pH meter (TOA HM-10P),
and conductivity meter (WTW LF 91), respec-
tively. Turbidity was measured using nephalome-
ter (ELICO, CL 52). The hardness, chloride, and
alkalinity were determined volumetrically. Fluo-
ride was measured by SPADNS method using acid
zirconyl–SPADNS reagent. Iron was estimated by
phenanthroline method (Greenberg et al. 1992).
In this method, the water samples (50 mL for
each) with 2 mL concentrated hydrochloric acid
and 1 mL hydroxylamine solution were heated
to boiling temperature in Erlenmeyer flask until
the volume reduces to 15–20 mL. The solution
was then cooled to room temperature and trans-
ferred into 50-mL volumetric flask with 10 mL
ammonium acetate buffer and 1 mL phenanthro-
line solution. The volume of solution in the flask
was maintained to the mark by adding de-ionized
water and placed for 10–15 min at room tempera-
ture. Absorbance of the sample was measured in
a colorimeter (CL 157, ELICO) at 510 nm wave-
length. The reference against de-ionized water at
480
zero absorbance was also measured to estimate
the total iron content by plotting a standard curve
with blank.
Arsenic was determined by using standard test
kit. Measuring efficiency of test kit was 0–0.5 mg/L
for arsenite (As, III) and arsenate (As, V). The
sample (20 mL) was placed in the standard size
reaction vessel. Reagents: arsenic 1 and arsenic
2 were simultaneously added into the reaction
vessel following the procedures mentioned in the
test kit manual. Test strip (∼2 cm) was inserted
into the reaction vessel and tightly clamped by
lid to avoid the leakage of arsine gas generated
inside the test vessel. Pressure balance inside the
vessel was adjusted by pricking the lid using sharp
hollow needle. The reaction vessel was then kept
for 30 min by gently swirling in between the
reaction time. After the reaction, the test strip
was carefully removed from the reaction vessel,
dipped into distilled water for 2 min and compared
the color developed in the test strip with standard
color scale in the manual.
Total coliform (TC) bacteria were quantified
by membrane filtration method (Greenberg et al.
1992). The samples (100 mL for each) were
filtered using sterile filter paper with pore size
of 0.45 μm by applying vacuum suction and in-
cubated in an incubator at 37◦ C for 24–48 h in
M-endo agar media. After incubation, total col-
iform bacteria were enumerated by counting the
colonies.
Results and discussion
A total of 87 groundwater samples were collected
from shallow wells (56), tube wells (20), and deep-
tube wells (11). The sampling sites and number of
samples collected from different sources are illus-
Table 1 Physical analysis of groundwater samples
Groundwater source Temperature (◦ C) pH
Min Max Aver Min
Shallow well (56) 12.8 22.0 18.6 5.9
Tube well (20) 11.2 23.1 17.9 6.7
Deep tube well (11) 14.3 24.7 20.3 6.6
WHO guidelines NA
NA not available
Environ Monit Assess (2011) 178:477–485
trated in Fig. 1. Of these groundwaters, shallow
wells are considered the crucial source of water
for domestic and other applications. People prefer
to dig shallow well rather than tube well and
deep-tube well because of less construction and
handling cost. The maximum number of samples
(56 samples) collected from shallow wells alone in
this work is apparent evidence to the importance
of shallow wells in the study area.
Physical and chemical analysis
The samples were analyzed for physical, chemical,
and microbiological parameters and presented the
results in average values. Physical analysis was
carried out to determine temperature, pH, EC,
and turbidity. The results are presented in Table 1.
EC of groundwaters were found to be 874.5,
576.8, and 704.2 μS/cm, respectively, for shal-
low well, tube well, and deep-tube wells whereas
turbidity was estimated to be 45.9, 54.8, and
33.2 NTU. These values are far above the limit of
WHO recommendations for drinking-water qual-
ity. High EC of water is due to the presence of
dissolved ionic substances, as EC increases with
increase in the ionic concentration. Different ionic
substances such as metallic ions, organic matters
containing ionic charges, ammonium, and nitrate
ions are some key parameters responsible for the
increase of EC of water. A similar study carried
out (Dzwairo et al. 2006) to determine the EC
of water also suggests that EC is due to ionic
substances and increases with the concentration of
ionic species in water. These substances are avail-
able to groundwaters by infiltration of agricultural
wastes, leakage of safety tank and also due to poor
sanitation of groundwater reservoirs. Maximum
EC of shallow water is due to easy availability
of such ionic substances to groundwater table.
Conductivity (μS/cm) Turbidity (NTU)
Max Aver Min Max Aver Min Max Aver
11.5 7.1 7.5 2640 874.5 0.4 750 45.9
7.7 7.0 345 974 576.8 0.4 420 54.8
7.5 7.0 286 2590 704.2 8.0 100 33.2
6.5–8.5 500 5
Environ Monit Assess (2011) 178:477–485 481

Aver
0.43
0.27
0.74
These materials get access to groundwaters either

Fluoride (mg/L)
by surface runoff or leaching from contaminated
sites and geological sources. In deep aquifer, the

Max
1.92
0.53
1.76
1.5
possibility of groundwater contamination is less
than that of shallow well because of the increased

Min
0.06
0.07
0.13
depth and safely covered top. The EC observed
in tube well and deep-tube wells may also be

0.004
0.003
0.009
Aver
primarily due to increased concentration of metal-
lic ions released from natural sources in addition

Arsenic (mg/L)
to anthropogenic activities. Turbidity of water is

0.025
0.010
0.025
Max

0.01
due to suspended matters derived from inorganic,
organic and microbial substances (Sawyer et al.

Min
2003). Increased turbidity of groundwaters, partic-

0
0
0
ularly in shallow water could be due to loose soil

Aver
structure around the sources along with the pres-

1.47
1.90
1.80
ence of suspended matters. As a consequence, the
soil gets disturbed and resuspend within the well

Max
Iron (mg/L)

5.5

0.3
4.9
5.2
during water withdrawal, increasing the turbidity
of water to a maximum degree. Other reasons of

Min
0.10
0.10
0.12
utmost turbidity in shallow water is poor main-
tenance and sanitation of groundwater sources

366.0
258.0
302.7
and improper construction pattern of safety tanks,

Aver
Alkalinity (mg/L)
which were very close to the groundwater sources
in almost all the places. Due to such conditions,

1000
650
550
NA
Max
suspended matters and microbial contaminants
can be easily available to groundwaters by leach- Min
ing through soil pores and leads to maximum tur-
100
100
50
bidity of water. In tube well and deep-tube wells,
it is less than that of shallow wells. The reason of
Aver
81.8
61.1
59.0

less turbidity may be due to properly covered top

Chloride (mg/L)

and increased depth of groundwater table.

198.2
127.8
383.4
Max

The pH was within the recommendations of

250

WHO drinking-water quality guidelines and ob-

served at neutral pH value (pH 7). The tem-
Min
4.26
14.2
7.10
Table 2 Chemical analysis of groundwater sample

perature of groundwaters was measured to be

18–20◦ C. There is no WHO guideline value for
230.7
218.8
251.2
Aver

drinking water temperature and also has not

Hardness (mg/L)

any significant importance from health point of

Max

view. However, the European Economic Commu-

800
500
640
500

nity and Canadian drinking-water guidelines have

recommended maximum drinking-water temper-
Min
050
130
078

atures of 25◦ C and 15◦ C, respectively (National

water quality management strategy 1996).
Groundwater source

Deep tube well (11)

Chemical parameters were estimated to deter-

Shallow well (56)

NA not available
WHO guidelines

mine hardness, chloride, alkalinity, iron, arsenic,

Tube well (20)

and fluoride in groundwaters (Table 2). Of these

parameters, hardness and chloride concentration
satisfied the WHO drinking-water quality recom-
mendations (Fig. 2).
482
Fig. 2 Mean concentration (±SD) of hardness, chloride,
and alkalinity in groundwaters
Hardness of water is due to presence of soluble
metal salts, particularly the calcium and magne-
sium carbonates. Hard water has not any negative
health impact; however, the use of hard water for
industrial applications especially for laundry ope-
ration cannot be recommended because of lea-
thering problem. Minimum amount of chloride
estimated in groundwaters suggest that the chloride
rich minerals are not present in the groundwater
table of Kathmandu valley. Estimated chloride
may be mostly derived from domestic effluents,
fertilizers, septic tanks, and natural sources such
as rainfall and weathering of chloride bearing min-
erals (Ritzi et al. 1993; Hem 1992). The alkalinity
Fig. 3 Mean concentration (±SD) of iron, fluoride a, and arsenic b in groundwaters
Environ Monit Assess (2011) 178:477–485
of groundwaters was 366, 258, and 302.7 mg/L,
in shallow well, tube well and deep-tube wells,
respectively. Alkalinity is a measure of its capacity
to neutralize acids and is primarily due to presence
of the salts of weak acids especially the carbonate
and bicarbonates. The ammonia or hydroxides
may also contribute total alkalinity of water. Max-
imum alkalinity in shallow water is probably due
to infiltration of sewage from safety tank and also
due to agricultural runoff, in addition to natural
sources. The alkalinity of groundwaters decreased
to lower level in tube well and deep-tube wells.
This variation in alkalinity reveals that the natural
factors responsible for the increment of alkalinity
are not prominent in groundwater table. There
is no standard guideline value for alkalinity of
drinking water and also has little public health
significance.
Iron concentration in groundwaters exceeded
the maximum limit of WHO drinking-water qual-
ity guidelines (Fig. 3a). Maximum iron (1.90 mg/L)
was estimated in the samples collected from tube
well and deep-tube wells. The iron concentration
in shallow water (shallow well) was 1.47 mg/L,
which is minimum amount of iron estimated in
groundwaters. Maximum iron in deep aquifer is
possibly due to increased amount of reduced iron.
The reduction of iron increases with increase of
the depth of groundwaters because of less avail-
ability of oxygen in deep aquifer. In general, iron
exists in water in soluble ferrous state, and can re-
main in the solution only in the absence of oxygen.
Environ Monit Assess (2011) 178:477–485
When the water containing ferrous ions comes
in contact with oxygen, the soluble ferrous ions
oxidize to insoluble ferric state and aggregate in
the form of precipitate. In the presence of oxygen,
the soluble ferrous ions in the water precipitate
into insoluble ferric states of geothite (FeOOH)
or ferrihydrites [Fe(OH)3 ], as illustrated in the
following chemical equations (Komnitsas et al.
2004; Houben 2003a, b).
2Fe2+ + O2 + 2OH− −→ 2FeOOH ↓ (1)
4Fe2+ + O2 + 8OH− + 2H2 O −→ 4Fe(OH)3 ↓
(2)
This condition greatly affects to the availability
of iron in groundwaters. The groundwater is gen-
erally deficit of oxygen and oxygen concentration
gradually decreases with increase of the depth
of groundwater table, resulting in increase of the
concentration of reduced iron in deep aquifer.
In shallow wells, some of the iron precipitates
into insoluble ferric oxide due to natural aeration,
resulting in reduction of the iron concentration in
shallow water. Iron is an essential metal required
in small amount by all living organism (Crichton
1991). Nevertheless, excess iron causes toxicolog-
ical problems in terms of accidental acute expo-
sures and chronic iron overload. Ingestion of iron
in excess amount (>0.5 g) causes severe toxicity
in human physiological systems leading to liver,
heart and lung diseases as well as diabetes mel-
litus, hormonal abnormalities, and dysfunctional
immune system (Gurzau et al. 2003). High con-
centration of iron makes water unusable due to
discoloration, metallic taste and odor, turbidity,
staining of laundry, and plumbing fixtures. More-
over, the growth of iron bacteria in iron containing
water causes corrosion in water supply pipes and
produce bad odors and unpleasant tastes in water
(CDPHE 2009; Kontari 1988).
Arsenic and fluoride concentration satisfied the
recommendations of WHO drinking-water qual-
ity guidelines. As depicted in Fig. 3b, maximum
arsenic estimated in groundwaters was 0.003–
0.009 mg/L. This is the acceptable concentra-
tion of arsenic recommended by WHO. Arsenic
483
is available to groundwaters either by smelting
of arsenic containing minerals or as a result of
improper agricultural practices and unsystematic
industrial activities. In this work, minimum con-
centration of arsenic estimated in groundwaters
reveals that the arsenic containing sources in
the groundwater aquifer of Kathmandu valley
is very low. Fluoride was estimated 0.43, 0.27,
and 0.74 mg/L in shallow well, tube well, and
deep-tube wells, respectively. These values satisfy
the WHO guidelines. Fluoride in groundwaters
is mostly of geological origin (Luna and Melian
2003).
Microbiological analysis
TC bacteria in groundwaters extremely exceeded
the WHO guidelines (Table 3). Bacterial popu-
lation ranged 0 to 300, with 8% of the total re-
sults meeting the 0 CFU/100 mL WHO guidelines
value. The average value of TC bacteria varied
in groundwaters and maximum coliforms were
enumerated in the samples collected from shal-
low wells (267 CFU/100 mL), while in tube well
and deep-tube wells, coliform count was 129 and
148 CFU/100 mL, respectively (Fig. 4).
Maximum coliform detected in shallow water
is probably due to poor drainage facility and im-
proper construction pattern of septic reservoirs.
Construction of septic tank close to the ground-
water sources may be a reason of high microbial
contamination to groundwaters, which was com-
mon almost in all the places as observed. As a
consequence, the effluent from septic tank can
easily percolate down to groundwaters and leads
high microbial contamination to groundwater ta-
ble. In tube well and deep-tube wells, TC count
was less because of increased depth and properly
covered top of groundwater sources, which was
not common in shallow wells. Another reason of
Table 3 Microbiological analysis of groundwater samples
Groundwater source Total coliform/100 mL of water
Minimum Maximum Average
Shallow well (56) 0 300 267.2
Tube well (20) 0 300 129.1
Deep tube well (11) 0 300 148.1
WHO guideline 0
484
Fig. 4 Mean value of total coliform bacteria (±SD) in
groundwaters
microbial contamination to groundwaters may be
due to direct discharge of untreated sewage or
municipal wastes into surface waters or in open
places near to groundwater sources, which was ob-
served in most of the places. Due to such unusual
practices, contaminants can easily leach down to
groundwater table leading high microbial conta-
mination to shallow water.
These results are obviously stating that the
groundwaters of Kathmandu valley contain excess
amount of iron and coliform bacteria. In addition
to iron and coliform, nitrogen containing sub-
stances (ammonia, nitrate, and nitrite) and pes-
ticide residues could also be present in ground-
waters of Kathmandu valley. These substances
further degrade water quality to a lower level
and make the water unacceptable for drinking
purpose. Sources of these contaminants may be
nitrogenous fertilizers and pesticides, which are
frequently used in agricultural fields to increase
crop productivity. Khadka (1993) and Krapac
et al. (2002) also suggests that nitrogen based
compounds such as nitrate, nitrite, and ammonia
are basically derived from chemical fertilizers and
domestic effluent. These pollutants accessed to
groundwaters by infiltration through soil pores,
leading groundwater pollution to a maximum de-
gree. The groundwaters of Kathmandu valley at
present cannot be recommended for drinking pur-
pose because of excess iron and coliform bacteria.
Environ Monit Assess (2011) 178:477–485
Conclusions
Groundwater quality of Kathmandu valley was
studied and assessed the contaminants level in
groundwater aquifer. The groundwater samples
were collected from shallow well, tube well,
and deep-tube wells located at different places
of Kathmandu valley. The physical, chemical,
and microbiological parameters of the samples
were evaluated to investigate the groundwater
quality for drinking water. The groundwaters
in the valley were found to be contaminated
with iron and coliform bacteria. A maximum
of 1.9 mg/L iron was estimated in the sam-
ples of tube well and deep-tube wells. Iron con-
centration in shallow water was approximately
1.5 mg/L. These values are well above the rec-
ommendations of WHO drinking-water quality
guidelines (0.3 mg/L). Total Coliform bacteria
enumerated in shallow wells greatly exceeded the
WHO guidelines (267 CFU/100 mL). Bacterial
population in tube well and deep-tube wells was
129 and 148 CFU/100 mL, respectively. Besides
iron and coliform bacteria; ammonia, nitrate, ni-
trite and pesticide residues also negatively im-
pact on groundwater quality. Physical parameters
estimated for electrical conductivity and turbid-
ity exceeded upper limit of the WHO standard
for drinking water. Hardness, chloride, arsenic,
and fluoride concentrations were within the lim-
its of WHO recommendations for drinking water
standard.
Recommendations
Ground water quality of Kathmandu valley is
poor due to presence of excess iron and col-
iform bacteria. The iron and coliform containing
water is suggested for adequate treatment before
use. Presence of maximum coliform bacteria in
groundwaters is mainly due to poor maintenance
of groundwater reservoirs and improper discharge
practices of household wastes into surface water
or in open places near the groundwater sources.
For the protection of groundwaters from bacterial
contamination, safe disposal of wastes by keep-
ing the groundwater reservoirs neat and clean is
recommended.
Environ Monit Assess (2011) 178:477–485
Acknowledgements This work was carried out under a
regular research program supported by Nepal Academy
of Science and Technology. The author is grateful to Dr.
Suresh Kumar Dhungel of Nepal Academy of Science and
Technology for proofreading of the manuscript. Thanks
to Gopal BK and Tista Prasai Joshi for their help during
sample collection and analytical work.
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