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 station.
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
Faridabad is 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.
The 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
7,083 km2 and passes a length of 400 km through Meerut District, Muzaffarnagar District, Ghaziabad,
Baghpat District, Noida, Greater Noida, prior to meeting Yamuna just exterior to Delhi.
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 Delhi.
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.
Table 7: Water Quality Samples Collected from Yamuna River, Hindon River,
Sewage water, Faridabad waste water, Canal water and Ground water.
Fig. 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 Sewage water
(c) Overnight culture on LB Agar plate of Ground Water
(d) Overnight culture on LB Agar plate of Hindon River
(e) Overnight culture on LB Agar plate of Faridabad waste water
(f) Overnight culture on LB Agar plate of Canal water
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 samples
(i) containing MacConkey Agar of Ground water samples
(j) containing MacConkey Agar of Hindon River samples
(k) containing MacConkey Agar of Faridabad waste water
(l) containing MacConkey Agar of Canal water samples
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 River samples
(h) were streaked on EMB Agar of Sewage water samples
(i) were streaked on EMB Agar of Ground water samples
(j) were streaked on EMB Agar of Hindon River samples
(k) were streaked on EMB Agar of Faridabad waste water
(l) were streaked on EMB Agar of Canal water samples
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 containing MacConkey
m, n, o, p, q & r : Pure colonies of E. coli lifted from g, h, i, j, k & l were
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
4.2 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.
All 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 d
e f g h
a: Indole c: VP
b: Methyl Red d: Citrate
e: Indole f: VP
g: Methyl Red h: Citrate
Fig. 13: (A) (a, b, c & d) Representative images used for biochemical