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(MPN/100ml)

: 500-Desirable2500-Max Permissible

2. Fecal streptococci(MPN/100ml)

: 100-Desirable500-Max Permissible

• Physical : Radiation – UV/ Cobalt

• Chemical : Chlorination, Ozonation, H2O2

• Biological : Bacteria in STPs

Sewage Treatment Plants(STPs) are Primarily designed for removal of BOD and SS but;

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 extra-cellular secretions

• Symbiotic : Law of limiting factor

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.

Lytic cycle:

a) Phage Adsorption on Host Cell.

b) Genetic Material Penetration of Phage.

c) Phage Multiplication.

d) Complete assemblage of Phages.

Life Cycle of Bacteriophage

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

Samples with Enrichment broth

Incubation

Isolation of Bacteriophage

Chloroform Precipitation, Centrifugation and Filtration

Mass Production

Shaking and Centrifugation

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/ InstrumentsIncubator Shake, High Speed Refrigerated Centrifuge, Filtration Assembly, Fermentors, Autoclaves (5 Sets for 5 Centres)

2 Glassware, Chemicals, Media, at 5 Centres

3 Staffing2 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

THANKSTHANKS