WATER QUALITY ASSESSMENT OF SELECTED

HOLY RIVERS POST GATHERING FOR THE

MAHAKUMBH 2025

Mentor- [Link] Mahanty

Research Associates - Khushi Dube, Aditi Jagtap, Khan Asahab,

Zeeshan Qureshi, Khushnuma Sheikh, Umera Datey

ACKNOWLEDGEMENTS
We would like to sincerely thank our mentor, Ms. Rituja Mahanty, for giving us the
opportunity to work on this project. Her guidance, encouragement, and patience meant a
lot to us throughout this journey. She has been a pillar of support throughout this project,
from conceptualization to execution, and we are truly thankful for her mentorship.

We would also like to extend our sincere thanks to all our teammates for their unwavering
cooperation, mutual respect, and the collaborative spirit that made this experience not only
productive but also enjoyable. Each team member played a vital role, and their dedication
and enthusiasm significantly contributed to the outcome of this project.

We’re also grateful to Jyoti Ma’am for her help and support whenever we needed it, and to
the Department of Biotechnology for providing us with the necessary resources and lab
facilities that helped us carry out our experiments smoothly.

Finally, we’d like to thank our college for giving us this opportunity to work on a hands-on
water quality analysis project. This project not only enriched our academic learning but also
gave us practical exposure and a deeper understanding of environmental and biological
aspects relevant to our field of study.

ABSTRACT
The Maha Kumbh Mela, a large-scale religious pilgrimage, increases anthropogenic pressure
on India’s sacred rivers, raising serious concerns about water quality and public health. This
study investigates the post-festival impacts on water samples collected from nine culturally
significant river sites, including the Ganga, Yamuna, Triveni Sangam (Prayagraj), Shipra
(Ujjain), Narmada, and the Narmada-Kaveri confluence. Samples were analysed for physical
and biochemical parameters like pH and IMViC, respectively, faecal coliform counts using the
Most Probable Number (MPN) method, and antimicrobial activity using Agar Well Diffusion
against Staphylococcus aureus and Pseudomonas aeruginosa. This study aims to assess the
impact of mass gatherings on river pollution and to explore the scientific validity of cultural
beliefs that associate river water with the healing of various ailments, including skin diseases
and gastrointestinal disorders. The MPN test confirmed the presence of faecal coliforms,
indicating microbial contamination. Antimicrobial Susceptibility Test revealed no zones of
inhibition, suggesting the absence of antimicrobial activity in the water. The pH levels ranged
from 7.0 to 8.5, indicating neutral to slightly alkaline conditions. These results underline the
need for continuous water quality monitoring during mass pilgrimage events. While the faith
attributes therapeutic effects to these waters, microbial risks persist.

INTRODUCTION
India’s rivers are far more than just natural water bodies — they are sacred lifelines that
nourish ecosystems, support livelihoods, and serve as the spiritual backbone of the nation.
For centuries, these rivers have been revered by millions, forming an essential part of daily
life, religious practices, and cultural identity. Rivers like the Ganga, Yamuna, Godavari, and
Narmada are not only sources of drinking water and irrigation but are also considered holy
by devotees who gather in large numbers to perform rituals. According to the Central
Pollution Control Board (CPCB, 2022), there are currently 311 identified polluted river
stretches across India. Many of these are contaminated with high levels of biological
pollutants, including coliform bacteria and pathogenic microorganisms, which are known to
cause waterborne diseases. These contaminants are particularly concerning in regions with
heavy human activity and poor sanitation infrastructure.

Large-scale religious events, such as the Kumbh Mela that recently took place, further
intensify the burden on these already stressed rivers. During such events, millions of pilgrims
gather along the riverbanks to perform ritual baths, which are believed to cleanse sins and
have healing properties. These rituals often involve drinking the river water, bathing, and
performing ceremonies involving offerings. While being spiritually significant, these activities
result in a dramatic increase in organic matter and microbial load in the water. Offerings such
as ashes of the deceased, idols, flowers, and coins are commonly deposited into the river,
often wrapped in polythene bags. Among the various indicators of river pollution, faecal
contamination stands out as a critical parameter. It reflects the presence of enteric
pathogens, including bacteria like E. coli, which can lead to outbreaks of gastrointestinal
diseases. The detection of such pathogens is a clear signal that the water is unsafe for
human use without proper treatment. In light of these concerns, the present study aims to
investigate the level of microbial contamination (focused on coliforms only) in selected holy
rivers after major religious festivals. Alongside this, the research also explores the
antimicrobial properties of the water — a concept rooted in traditional beliefs that such
water from the significant holy rivers, possesses healing and purifying qualities.

MATERIALS AND METHODS

Sample collection:

A total of nine water samples were collected from key locations along the banks of some of
India’s most prominent rivers, namely the Ganga, Yamuna, Narmada, Shipra, Triveni Sangam,
and a confluence of Narmada and Kaveri. These sites were chosen due to their high religious
significance and the large number of people who gather there for rituals, particularly during
festivals and special occasions. Immediately after collection, the samples were labelled.
Upon arrival at the laboratory, the samples were refrigerated at appropriate temperatures to
preserve their physical and biological characteristics until further analysis.

Most Probable Number (MPN):

This test was conducted in three steps as follows:

1) Presumptive test
2) Confirmatory test
3) Complete test

Presumptive test: This is the primary presumptive analysis for to check for the presence of
Coliforms in the samples. In this test, Teepol broth with durham’s tube was used as teepol
broth contains lactose as one of its constituents and is fermented by the coliforms resulting
in a colour change from reddish to yellow with a simultaneous release of gas detected by the
collection of gas in the durham’s tube. A set of 5 tubes were used for Double Strength broth
+ 10 ml sample for each water sample, a set of 5 tubes used for Single Strength broth + 1 ml
sample for each water sample and a set of 5 tubes used for Single Strength broth + 0.1 ml
sample for each water sample. These were further incubated at 37˚C for 48 hours. The
results were compared to a standard MPN chart for presumptive coliform count per 100 ml
of water sample.

Confirmatory test: The tubes showing positive results from the presumptive step were
selected and streaked on Eosin Methylene Blue (EMB) Agar. This further distinguishes
organisms, E. coli shows a metallic sheen on EMB agar whereas other gram-negative
coliforms show black centred colonies after incubating the plates at 37˚C for 24 hours.

Complete test: The bacterial colonies that indicated presence of coliforms were further
inoculated in Lactose Bile broth with a durham’s tube in it. The colour change after
incubation at 37˚C for 48 hours in the lactose bile broth along with the presence of gases in
the durham’s tube confirms the presence of coliforms. For further compete confirmation,
Indole, Methyl red, Voges-proskauer and Citrate (IMViC) and Gram staining was done to
characterize the coliforms even further.

Antibiotic Susceptibility Test (AST):

It was performed using the agar well diffusion method to evaluate the potential
antimicrobial or healing properties of the water samples, as various religious beliefs support.
The test organisms, Pseudomonas aeruginosa and Staphylococcus aureus, were selected as
representative Gramnegative and Gram-positive bacterial strains, respectively. Soft nutrient
agar butts (20ml) were inoculated with these bacterial suspensions (1ml) and poured onto
the petri plates, and wells were prepared on these plates into which the river water samples
were aseptically introduced. Ampicillin was taken as a positive control, and distilled water as
a negative control. Plates were incubated at 37ºc, and the presence or absence of a zone of
inhibition around each well was recorded after the incubation period.

RESULTS AND DISCUSSION

1) Most Probable Number (MPN) Test

It was observed that all the 9 samples showed some levels of lactose fermentative bacteria
(coliforms) as the result ranged from 4 to ≥1600 MPN/100 ml of sample. (see figure 1 and 6)
The confirmatory test showed 5 out of the 9 samples to be contaminated by gram negative
coliforms. (see figure 2 and 8) And the complete test confirmed the presence of coliforms in
the 5 samples that were observed positive for the confirmatory test. (see figure 3 and 7)
Further IMViC and Gram staining characterized the organisms from these 5 samples. (see
figure 4,5,9 and 11)

2) Antibiotic Susceptibility Test

All tested river samples showed no zones of inhibition against both of the test organisms,
indicating a lack of detectable antimicrobial activity under the experimental conditions used.
(see figure 12)

Number of positive tubes in

Sample 10 ml 1 ml 0.1 ml MPN index
sample sample sample
Yamuna 1 1 0 4
Sample 9 2 0 0 4
Triveni Sangam 1 3 2 0 14
Triveni Sangam 2 4 1 0 17
Narmada 3 2 1 17
Narmada + Kaveri 5 5 0 240
Ganga 1 5 5 0 240
Ganga 2 5 5 3 900
Shipra 5 5 5 ≥1600
Figure 1: Presumptive test results for MPN

Sample Growth Colonies with Colonies with Result

observed metallic sheen black centre
Yamuna + - - Safe
Triveni Sangam 1 + - - Safe
Ganga 1 + - - Safe
Sample 9 + - - Safe
Narmada + - + Unsafe
Narmada + Kaveri + - + Unsafe
Triveni Sangam 2 + - + Unsafe
Ganga 2 + + - Unsafe
Shipra + - + Unsafe
Figure 2: Confirmatory test results for MPN

Sample Colour change of LB Gas released Results

 broth
Narmada + + Unsafe
Narmada + Kaveri + + Unsafe
Triveni Sangam 2 + + Unsafe
Shipra + + Unsafe
Ganga 2 + + Unsafe
Figure 3: Complete test results for MPN

Sample Indole Methylred Voges- Citrate Interpretation

Proskauer
Narmada + + + + Klebsiella oxytoca
Narmada + Kaveri - - + + Klebsiella pneumoniae
Triveni Sangam 2 - - + + Klebsiella pneumoniae
Shipra + + - - E. coli
Ganga 2 + + - + Citrobacter koseri
Figure 4: Characterization of coliforms in positive samples using IMViC

Sample Gram Nature Colony Characteristics

Narmada Negative Mucoid and raised colonies
Narmada + Kaveri Negative Mucoid, and sticky colonies
Triveni Sangam 2 Negative Mucoid, and sticky colonies
Shipra Negative Medium-sized, smooth, round colonies
Ganga 2 Negative Round, moist colonies; slightly raised
Figure 5: Characterization of coliforms in positive samples using Gram staining
Figure 6: Presumptive test for MPN

Figure7: Complete test for MPN

Figure 8: Confirmatory test for MPN

 Figure 9: IMViC for further characterization

Figure 11: Gram staining for further characterization

 Figure 12: Antibiotic susceptibility test

CONCLUSION
This study highlights the presence of coliforms in four out of nine river samples (Narmada,
Triveni Sangam, Ganga, Narmada and Kaveri confluence) and faecal coliforms in the River
Shipra sample collected post the Maha Kumbh gathering, indicating potential health hazards
for communities relying on these waters. However, the number of samples analysed were
relatively limited when compared to the scale of a full-scale environmental project. Hence,
future studies must involve a larger sample size to gain a more detailed understanding,
particularly focusing on faecal contamination levels. To improve water safety of such
contaminated river water can be treated through methods such as chlorination, UV
disinfection, membrane filtration, constructed wetlands, and biological treatment systems.
Continued monitoring and corrective actions are essential for preserving river health and
public well-being.