Transcript Elimination of escherichia coli and other coliform bacteria through

Central Pollution Control Board

29 cities, 70 towns and many villages:

Sewage disposal-a major cause of fecal coliform

Disposal of 3000 MLD sewage and having 1000
MLD treatment capacity only.

Exceedance of Fecal coliform in Gaga

Fecal coliform the cause of concern.

Fecal coliform converts water unfit for Bathing
and for other designated best uses.
1.
Fecal coliform
:
500-Desirable
2500-Max Permissible
:
100-Desirable
500-Max Permissible
(MPN/100ml)
2.
Fecal streptococci
(MPN/100ml)
• Physical
:
Radiation – UV/ Cobalt
• Chemical
:
Chlorination, Ozonation, H2O2
• Biological
:
Bacteria in STPs

Sewage Treatment Plants(STPs) are
designed for removal of BOD and SS but;
Primarily

Not designed specifically to treat fecal coliform.

Bactericidal actions like chlorination, UV radiation,
Ozonation and other oxidants are employed.
[Irradiation generates Mutants of un-known attributes]

Antagonists Bacteria

Bacteriophage and their synergy
Biological Action
• Competitive
:
Competition for food
• Inhibitive
:
Lethality
of
secretions
• Symbiotic
:
Law of limiting factor
extra-cellular
The consortia acts on three mode;
Competitive: Certain microbes grow and multiply at a faster rate and
create competition for food. Dominant forms survive
and others become recessive/ dormant.
Inhibitive
: Some forms secretes extra-cellular enzyme becoming
lethal to other form of bacteria and act as antagonists.
Symbiotic : In a symbiotic actions, Bacterial forms grow together on
a common food plank (substrate) helping each other to
grow but restrict growth of other forms due to law of
limiting factor.
Life Cycle of Bacteriophage
Lytic cycle:
a) Phage Adsorption on Host Cell.
b) Genetic Material Penetration of Phage.
c) Phage Multiplication.
d) Complete assemblage of Phages.
Lysogenic cycle:
The genetic material of Phage remains within the host cells and
reproduce in synchrony with the host for long-period.
Lysogenic and Lytic cycle
Isolation of Bacteriophage
Samples with Enrichment
broth
Incubation
Shaking and Centrifugation
Chloroform Precipitation,
Centrifugation and Filtration
Mass Production
Sewage/ Sludge
(Broth Culture)
Incubation
Shaking Incubation
Centrifugation
Rinsate Centrifuge
Shaker incubator
Chloroform – Enriched single type of
phage
Lyophilised

The concentration of targeted bacteria Vs the
concentration of phages.

The phages will be introduced into the STP/ drain
either batch wise or in continuous flow and testing
to determine reduction of targeted bacteria.
Application of Mass culture with pre-determined doses for
removal of E.coli and other fecal coliform;

Sewage Treatment Plants established under GAP

Sewage drains joining river Ganga
Phage treatment demonstration;
Installed STPs and Drains carrying sewage joining river Ganga
in; Uttarakhand, Uttar Pradesh, Bihar and West Bengal.
Duration of Project : 18 months
Mass Culture (BP) and experimentation on :
• Central Laboratory
:
CPCB, Delhi
• Uttarakhand
:
(SPCB)
• Uttar Pradesh
:
(Zonal Office of CPCB and SPCB at Lucknow/
Kanpur)
• Bihar
:
SPCB (Patna)
• West Bengal
:
Zonal Office of CPCB at Kolkata and SPCB
1
Infrastructure/ Instruments
Incubator Shake, High Speed Refrigerated Centrifuge, Filtration Assembly,
Fermentors, Autoclaves (5 Sets for 5 Centres)
2
Glassware, Chemicals, Media, at 5 Centres
3
Staffing
2 RA, 2 JRF and 2 PA at each Centre (5 Centres)
(to be co-ordinated by one Scientist ‘B’/ ‘C’ at each Centre)
4
Experimentation-one drain and one STP in: Delhi, Uttarakhand, Uttar
Pradesh, Bihar and West Bengal (Includes Phage dosing devices, minor Civil
works, performance/ testing charges) estimated expenditure at each centre
will be ` 1.0 Crore
5
Contingencies, Project expenses TA/DA, Meetings,Awareness
1 Procurement
:
3 months
2 Recruitment of Staff
:
3 months
3 Experimentation
:
12 months-culturing,
Application and
Performance monitoring
Studies
I
Instruments
(i)
High Speed Refrigerated Centrifuge
(ii)
Incubator Shaker
(iii)
Autoclave
(iv)
Filtration Assembly
(v)
Fermentors (10 L/20 L)
(vi)
Laminar flow
II
Glass Wares, Chemical, Media and
Lyophilisation Cost (per Centre)
III
Staffing (per Centre)
(i)
Field Labour Hiring
IV
Experimentation
Dozing and mixing devices (Filters, Blocks, Arms)
for BP on Sewage in STP and Drain (5 Centres)
V
Contingencies (institutional Charges, Awareness,
Travel and others)
Phage-therapy will be promising for us
THANKS