4.1 Collection of water samples and isolation
of the E. coli & K. pneumonia
The water samples collected from
the different ecologically important locations in Delhi. They are seven
different sites of Yamuna river, three sites of sewage water, canal water as well as Hindon river, Faridabad
waste water and two sites of Ground water. Further details of the sites and the
sample collection methods are described in materials and method). Each of these
collected samples were subsequently streaked onto Petri dishes containing suitable
growth medium. The initial screening of E.
coli & K. pneumoniae was done
Eosin Methyl Blue Agar (EMB) plates. A total of Ninety six E. coli & Eighty-four K.
pneumonia isolated were biochemically
identified collected from different geographical sites.
The water of Yamuna is of “reasonably good quality”
through its length from Yamunotri in the Himalayas to Wazirabad in Delhi, about
375 kilometers, where the discharge of waste water through 15 drains between
Wazirabad barrage and Okhla barrage renders the river severely polluted after
Wazirabad. It’s located in west Delhi.
ISBT (Kashmiri Gate)
About 4.4 km from Wazirabad
barrage near ISBT Kashmere Gate, close to the Metro Rail Power feeder
Sarai Kale Khan
14 km downstream from Wazirabad
barrage (near Sarai Kale Khan bus stand) at Delhi – Ghaziabad (Noida) road
bridge on Yamuna River. The water quality at this location reflects the
impact of wastewater, agriculture and animals discharge.
Rajghat is a memorial dedicated to Mahatma Gandhi. Originally it
was the name of a historic ghat of Old
Delhi (Shahjahanabad) on the banks of Yamuna river. Close to it, and east of Daryaganj was Rajghat
Gate of the walled city, opening at Rajghat on Yamuna River.
Later the memorial area was also called Rajghat.
Faridabad Waste Water
the largest city in the north Indian state
of Haryana in Faridabad district. It is a leading industrial
centre and situated in the National Capital Region bordering the
Indian capital New Delhi. Located 284 kilometers south of the state
capital Chandigarh, it is the most populous city in the state
of Haryana. The river Yamuna forms the eastern district
boundary with Uttar Pradesh.
24 km downstream from Wazirabad
barrage and East of Delhi-Agra National highway near Madanpur Khadar village
in South Delhi. The water quality at this location reflects the impact of
discharge of treated, partially treated effluents from Okhla Sewage treatment
Plant, other drains joining the river/canal and Hindon-cut.
Canal is located at the North
part of Delhi connecting Haidarpur water Treatment Plant to Hariyana.
Drinking water supplied to Haidarpur plant from Haryana for local water
supply in Rohini.
The DND Flyway
(Delhi Noida Direct Flyway) is an eight-laned 9.2 km access
controlled tolled expressway which connects
Delhi to Noida, an industrial suburb area. It was built
and is maintained by the Noida Toll Bridge Company Ltd promoted the
Infrastructure Leasing and Financial Services (IL).The project
included the construction of a flyover at Ashram Chowk which was handed over
to the Public Works Department for maintenance. A 552.5 meter bridge over the Yamuna river was
constructed at a cost of ?4.08 billion (US$63.6 million) and
was completed by February 2001.
Hindon River is a wholly rain-fed river. The river has its source in the
Saharanpur District from Upper Shivalik in lower Himalayan Range. The Hindon
River has a drainage basin of
km2 and passes a length of 400 km through Meerut District, Muzaffarnagar District,
District, Noida, Greater Noida, prior to meeting Yamuna just exterior to
This particular drainage is a
very important part of wastage of North Delhi covering entire rural part,
Rohini and Shalimar Bagh area of North Delhi. This is one of the polluted
drain in North Delhi.
Punjabi Bagh sewage
This particular drainage is a
very important part of wastage of West Delhi, covering Punjabi Bagh, Moti
nagar, Rajouri garden and Pashchim vihar area. This is one of the polluted
drain in west Delhi.
Hasanpur village sewage
This particular drainage is a
very important part of wastage of East Delhi, covering entire shahdra,
Krishna nagar and Patpadganj area. This is one of the polluted drain in East
Yamuna Pushta is the Pushta (embankment) on both sides of
the Yamuna River in Delhi, starting from the ITO bridge and up to the Salimgarh
Fort. It has also been home to riverbed
cultivators, and over 100,000 residents a string of slum colonies (shantytown) for some 40 years, mostly on the western banks, close
proximity of Old Delhi and a few on
the eastern banks like those near Sakarpur village in East Delhi.
7: Water Quality Samples Collected from Yamuna River, Hindon River,
Sewage water, Faridabad
waste water, Canal water and Ground water.
9: Overnight culture on LB Agar plate
(a) Overnight culture on LB Agar plate
of Yamuna River
(b) Overnight culture on LB Agar plate of
(c) Overnight culture on LB Agar plate of
(d) Overnight culture on LB Agar plate of
(e) Overnight culture on LB Agar plate of
Faridabad waste water
(f) Overnight culture on LB Agar plate of
Fig.10: Pure colonies lifted from a, b,
c, d, e & f were streaked on petriplate
(g) containing MacConkey
Agar of Yamuna River samples
(h) containing MacConkey Agar of Sewage water
containing MacConkey Agar of Ground water samples
containing MacConkey Agar of
Hindon River samples
containing MacConkey Agar of Faridabad waste water
(l) containing MacConkey Agar of Canal water
Fig. 11: Pure colonies of E. coli lifted from, g,
h, i, j, k & l containing MacConkey agar,
were streaked on petriplate shown in,
(g) were streaked on EMB Agar of Yamuna
were streaked on EMB Agar of Sewage water samples
(i) were streaked on EMB Agar of Ground water
(j) were streaked on EMB Agar of Hindon
(k) were streaked on EMB Agar of Faridabad
(l) were streaked on EMB Agar of Canal water
Fig. 12: Pure colonies of K. pneumonia lifted from B were streaked on EMB
a, b, c, d, e & f : Overnight culture
on LB Agar plate
g, h, i, j, k & l Pure colonies lifted
from a, b, c, d, e & f were
streaked on petri plate shown in g, h, i, j, k & l
m, n, o, p, q & r : Pure colonies of E. coli lifted from g, h, i, j, k &
streaked on EMB Agar
s, t, u, v, w & x : Pure
colonies of K. pneumonia lifted from
g, h, i , j, k &
l were streaked on EMB Agar
Antibiotic resistance offered by different environmental strains
In general, a bacterium goes through
four distinct phases; a lag phase, log phase, stationary
phase, and a death phase. The lag phase shows
how bacteria reproduce at a very slow
rate at first. At this point, the cells are
preparing for the division.
They are making sure to manufacture
fats and protein for the reproduction ahead. The second phase is the log (logarithmic or
exponential) phase. The bacteria is now replicating rapidly and becoming so
large in numbers that space is growing smaller, as is non-hazardous room and
nutrient. Due to this rapid growth, the next step is the stationary phase. In
this phase, about fifty percent of the bacteria population will become
inactive, and the other fifty percent will remain and continue replication
(binary fission). The last stage of bacteria generational grow is the death
stage. In the death stage, there are not enough nutrients for the entire
population. This causes the death rate of E.coli & K.
pneumonia to increase, and the division will slow as well. So, the growth
of microorganisms can be divided into 4 main phases: lag time (no growth),
exponential growth phase, stationary phase, and death phase.
4.2.1 Biochemical Assay for E. coli & K. pneumonia
For the confirmation of previously
identified and presumptive E. coli
& K. pneumonia strains,
biochemical assay was performed using four different tests for the confirmatory
identification of Escherichia coli & K. pneumonia. All 180 E. coli & K. pneumonia strains were checked by biochemical test (IMViC tests)
for reconfirmation of presence of E. coli
& K. Pneumonia. Details of the
biochemical characteristics of the E.
coli & K. pneumonia strains
which were investigated further are summarized in Table
A in Appendix 2 & Figure 13.
the 180 samples were confirmed by biochemical test and identified as E. coli & K. pneumonia.
Thus it was observed that 53.33% of
the E.coli & 46.66% of the K. Pneumonia isolated from the Yamuna
River, Hindon River, Sewage water, Ground water and other water bodies.
a b c
e f g
Indole c: VP
b: Methyl Red d: Citrate
Indole f: VP
g: Methyl Red h: Citrate