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