Determination of fecal indicator bacteria in

shallow and deep groundwater sources in the

Kathmandu valley, Nepal
D. Bhandari1*, S. Tandukar1, E. Haramoto2, J.B. Sherchand1
1Public

Health Research Laboratory, Tribhuvan University, Institute of Medicine, Nepal

Research Center for River Basin Environment, University of Yamanashi, Japan
*Corresponding author: me.dinesh43@gmail.com

2Interdisciplinary

Abstract Groundwater is a crucial source of water

for the residents of the Kathmandu valley, with an
estimated 50% of total inhabitants dependent on
groundwater supply for daily use. Hence, microbial
evaluation of groundwater is a paramount exigency to
evaluate the impact on public health of fecal
contamination of the groundwater. The purpose of this
study was to compare the distribution of fecal indicator
bacteria in both shallow and deep groundwater sources
of the Kathmandu valley as well as to evaluate their
microbial quality. A total of nine groundwater sources
within the Kathmandu valley including four shallow
water sources and five deep-bore wells were sampled
during the study period (May-July 2015) and tested for
Escherichia coli and total coliforms by the most
probable number (MPN) method using Colilert reagent
(Idexx Laboratories). Coliform bacteria were detected in
100% (9/9) of the groundwater samples tested and E. coli
was detected in 89% (8/9) of the samples tested. The
occurrence of E. coli in the shallow groundwater sources
was found to be 100% (4/4) and in the deep sources was
80% (4/5), with the average value ranging from 2 to
629.4 MPN/100 ml and <1 to 3 MPN/100 ml,
respectively. Similarly, the distribution of coliforms was
found to be 100% in both the shallow and deep
groundwater sources, with the average value ranging
from 960.6 to >2419.6 MPN/100ml and 18.7 to 155.3
MPN/100ml. respectively. Deep groundwater sources
were found to be comparatively less contaminated with
fecal indicator bacteria compared to shallow
groundwater sources. None of the groundwater samples
analyzed met the drinking water quality standard
recommended by World Health Organization, a finding
which warrants the implementation of an effective
treatment measure prior to use.
Keywords
Kathmandu

Groundwater,

Coliform,

E.

coli,

1. INTRODUCTION
Groundwater is a crucial source of water for the
residents of the Kathmandu valley. Currently,
approximately 50% of the water supply (estimated to be
59.06 million liters a day) that is used for drinking and other

domestic requirements in the Kathmandu valley are derived

from groundwater [1]. Several groundwater sources
including dug wells, shallow tube wells, stone sprouts and
deep tube wells are used in Kathmandu [2-3] with water
from the deep tube wells comprising about 32% (37.49
million liters a day) of the water used in the Kathmandu
valley [1].
Contextually relevant, recent studies have reported that
the groundwater in the Kathmandu valley has significant
microbial contamination that exceeds the drinking water
quality standard recommended by the World Health
Organization (WHO) guidelines [2, 4].
Waterborne pathogens are present in high concentration
in the feces of infected patients and they are transmitted
primarily through fecal-oral route, especially through
consumption of contaminated food and water [5]. In
addition to the risk of diarrhea from drinking water,
extensive use of groundwater for bathing purposes can be
implicated as an important transmission pathway through
accidental water ingestion leading to diarrhea [6].
Knowledge regarding the prevalence of waterborne
pathogens is essential to evaluate the risk of infection from
pathogens transmitted via contaminated water. Hence,
microbial evaluation of groundwater is a paramount
exigency to evaluate the impact of fecal contamination of
groundwater on public health. As revealed by this milieu,
the present study was undertaken to compare the
contamination intensity of shallow and deep groundwater
sources of the Kathmandu valley by fecal indicator bacteria.
2. MATERIAL AND METHODS
2.1 Study Site and Sample Collection
Three study sites in the Kathmandu valley,
(Kathmandu, Bhaktapur and Lalitpur), are located in the
upper part of the Bagmati River Basin in central Nepal. The
watershed has a total surface area of 665 km2. During the
months of May-July 2015 (i.e., mid-monsoon),
groundwater samples were collected from four shallow
groundwater sources [including two stone spouts located at
Sinamangal (Gw-Sw1), Kupandole-Lalitpur (Gw-Sw2) and
two shallow dug wells located at Nala-Bhaktapur (GwSw3), Kapan (Gw-Sw4)] and five deep groundwater
sources [including five deep bore wells located at
Sukedhara (Gw-Dw1), Gaushala (Gw-Dw2), Jagati-

Naresuan University Engineering Journal, Vol.11, Supplement, 1, May, pp 43-46

43

Bhaktapur (Gw-Dw3), Maitidevi (Gw-Dw4) and Sitapaila

(Gw-Dw5)]. The average depth of the shallow groundwater
sources ranged between 10 to 15 ft and those of the deep
groundwater sources ranged between 40 to 110 ft.
Water samples were collected in 100ml sterile
polyethene bottles and transported to the Public Health
Research Laboratory, Institute of Medicine within 2 hours
of collection for microbial analysis. The bottles were
thoroughly rinsed before sample collection.
2.2 Microbial Analysis
Total coliform and E. coli were measured by the most
probable number (MPN) method using Colilert reagent
(Idexx Laboratories) following the manufacturers
protocol. Briefly, 100 ml of sample water from a sterile
polythene bottle was thoroughly mixed with Colilert
powder and transferred in Quanti-tray. After removing the
bubbles inside the Quanit-tray, it was sealed using the
Quanti-tray sealer by placing the tray onto a Quanty tray
rubber insert of the sealer. Finally, the tray was incubated
overnight at 37 C and following the incubation, large and
small positive wells (yellow colored) were counted for total
coliform. For E. coli detection the tray was observed under
UVlamp and blue fluorescence in the wells was observed.
Ultimately, MPN was determined by referring to the
Quanti-tray MPN table.
3. RESULTS AND DISCUSSION
Total coliforms were detected in all nine groundwater
sample tested with an average value ranging from 960.6
to >2419.6 MPN/100 ml and 18.7 to 155.3 MPN/100 ml
respectively, in shallow and deep groundwater sources. E.
coli was detected in 89% (8/9) of the samples tested. The
occurrence of E. coli in the shallow and deep groundwater
sources was found to be 100% (4/4) and 80% (4/5), with an
average value ranging from 2 to 629.4 MPN/100 ml and <1
to 3 MPN/100 ml, respectively. Table 1 shows the results
of detection of total coliforms and E. coli in the shallow and
deep groundwater sources.
In the study, the entire groundwater samples tested were
found to be positive for total coliforms and 89% of the
sources were contaminated with E. coli. This compares
closely to several previous studies [4, 7, 8] in which severe
fecal contamination and microbial pollution of groundwater
in the Kathmandu valley have been documented. According
to the WHO drinking water guidelines, E. coli should not
be present in 100 ml of drinking water [9]. Out of the nine
groundwater samples tested, only one of the deep bore
wells in our study met the WHO drinking water guidelines
for E. coli.
Fecal pollution of groundwater can lead to health
problems because of the presence of infectious

44

microorganisms which may be derived from human sewage

or animal sources. The intestine of an infected vertebrate
may harbor various harmful microbes, including bacteria,
viruses, and protozoa, which may contaminate the water
source as indicated by the presence of fecal coliforms
derived from human sewage and animal droppings [10].
This is why coliform bacteria are considered indicator
organisms; their presence warns of the potential presence
of disease causing organisms. The E. coli test is especially
useful as an indicator organism because these microbes are
only related to fecal wastes. Therefore, while total
coliforms are a useful indicator of bacteria in the water,
E.coli is much more useful for determining whether water
has been contaminated from fecal wastes or not [15].
For the purpose of this study, groundwater sources (with
depth <20ft) were considered as shallow water sources and
those (with depth 20 ft) were considered as deep water
sources. Both the contamination intensity as well as the
burden of total coliforms and E. coli was found to be higher
in shallow water sources compared to deep water sources in
this study. Bacterial contamination of groundwater is
usually due to the mixing of surface runoff passing through
urban areas and pastures, leakage of sewage disposal
systems and septic tanks, overloaded sewage treatment
plants, disposal systems, and raw sewage deep well
injection [12]. In the Kathmandu valley, sewers are leaky
because of improper construction [13]. Most of the
contaminated water sources in our study were found to be
located closer to septic tanks, pit latrines and sewage drains.
Hence, it is likely that a huge amount of wastewater is
leaking into the subsurface of these water sources. In the
case of the Kathmandu valley storm drains and sewer lines
carry mixtures of sewage and storm water. The frequent
overflowing of such drains through manholes pollute the
land surface and subsequent seepage of the water from such
polluted surface results in contamination of subsurface
aquifers [14]. Likewise, infiltration of contaminants and
fecal materials generated from anthropogenic and zoonotic
sources may contribute in the contamination of shallow
groundwater sources [6].
In contrast to the shallow water sources, the deep
groundwater sources were found to be less contaminated
with both fecal coliforms and E. coli which is in accordance
with the findings of other studies conducted in Kathmandu
[5, 6]. However, total coliforms were detected in all the
deep water sources tested with an average value ranging
from 18.7 to 155.3 MPN/100 ml and E. coli was also
detected in 80% of the deep bore wells tested, although in
minimal quantity. Industrial and household waste is
discharged into the deep groundwater in the Kathmandu
valley [1], which could be one possible explanation for the
detection of total coliforms and E. coli in the deep water
sources tested.

Naresuan University Engineering Journal, Vol.11, Supplement, 1, May, pp 43-46

Table 1 The distribution of total coliforms and E. coli in the groundwater samples tested

No.

Sample ID

1
2
3
4
5
6
7
8
9

GW-SW1
GW-SW2
GW-SW3
GW-SW4
GW-DW1
GW-DW2
GW-DW3
GW-DW4
GW-DW5

Source type

Depth of the
source (ft)

Stone spout
Dug well
Dug well
Stone spout
Deep bore well
Deep bore well
Deep bore well
Deep bore well
Deep bore well

10
12
15
15
110
100
50
40
60

Location

Sinamangal
Nala
Kapan
Kupandole
Sukedhara
Gaushala
Jagati
Kupandole
Sitapaila

Distance
from septic
tank/sewage
drainage (m)
NA
20
8
6
5
NA
8
2
2

Total
coliforms
(MPN/100
ml)
1011.2
>2419.6
2419.6
960.6
79.4
18.7
36.4
101.2
155.3

E. coli
(MPN/100
ml)
629.4
16
2
23.5
<1
1
2
2
3

* NA- not available

4. CONCLUSIONS
This study found that deep groundwater sources of the
Kathmandu valley are less contaminated with fecal
indicator organisms than the shallow groundwater sources.
Most of the groundwater samples tested in the study were
found to be placed or constructed without fulfilling the
criteria laid down by the WHO which requires the source
of groundwater to be located more than 50 feet from likely
sources of contamination. This failure to observe the
criteria has led to the contamination of groundwater with
the fecal material due to seepage and percolation, resulting
in higher contamination of E. coli. Since more than 50% of
the population of the Kathmandu valley is dependent on
groundwater sources, the legal authority should facilitate an
effective treatment measure of the groundwater prior to use.
An immediate action with regard to preventing drinking
water sources from contamination is needed so as to prevent
the waterborne health problems and related morbidity and
mortalities in future
5. ACKNOWLEDGMENTS
The reagents and consumables used in this study were
supported by the Science and Technology Research
Partnership for Sustainable Development (SATREPS)
project `Hydro-microbiological approach for the water
security in Kathmandu Valley, Nepal`. . In addition, many
thanks to Mr. Roy Morien of the Naresuan University
Language Centre for his editing assistance and advice on
English expression in this manuscript.
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Naresuan University Engineering Journal, Vol.11, Supplement, 1, May, pp 43-46