En svensk vrektygslåda för fekal källspårning Dr. Jakob Ottoson, Assoc. Prof. Risk and benefit assessor, Microbiologist National Food Agency Dr.

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1 En svensk vrektygslåda för fekal källspårning Dr. Jakob Ottoson, Assoc. Prof. Risk and benefit assessor, Microbiologist National Food Agency Dr. Rikard Dryselius, Microbiologist

2 Safe water management WSP WPZ HACCP Know your catchment Inventory of pollution sources, effects of weather and other events. Know your source water Levels of indicators and their variation. Estimation of pathogen levels. Target your treatment Which barrier effect is needed with regard to water quality. Monitoring of treatment function. Protect your distribution system Pipe exchange rate, routines, protection of extraction wells, pressure in the pipeline.

3 Water safety planning Revise WSP following incident (Module 11) Assemble team (Module 1) Describe the water supply system (Module 2) incident Plan & carry out periodic WSP review (Module 10) Identify the hazards & assess the risks (Module 3) Develop supporting programmes (Module 9) Prepare management procedures (Module 8) Preparation System assessment Monitoring Management & communication Feedback Determine & validate control measures, reassess & prioritize risks (Module 4) Develop, implement & maintain an improvement plan (Module 5) Verify the effectiveness of the WSP (Module 7) Define monitoring of control measures (Module 6)

4 Protecting the health of Europeans by improving methods for the detection of pathogens in drinking water and water used for food production Aquavalens - consortium 39 partners 13 countries Universities, SMEs, Governmental agencies, Reasearch Institutes, Industry 9 million EU 12 million total University of East Anglia Protecting the health of Europeans against contaminated drinking water through increased understanding and accurate testing for waterborne pathogens Generating new knowledge on the molecular genetics of viral, bacterial and parasitic waterborne pathogens Developing new technologies to prevent and control the spread of waterborne disease

5 Aquavalens - research strategy 1.Applied microbiology: identification of molecular targets, markers and gene sequences providing a better insight in the occurrence of pathogens in water, their viability and infectivity. 2. Platforms for detection: development in order to enumerate these new sequences in water, automation, integration and standardisation of methods so they can be used in control programmes. 3. Test beds: design of sampling plans for large and small water companies and water for food production. 4. Effect, synthesising results to increase the knowledge on health risks with water, how these are affected by new pathogens and climate change. What effect has the implementation of water safety plans and how can new methods improve and help to enable the implementation?

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7 Utvärdering Aquavalens Fyra förslag (Blanch et al. 2015): Human vs. Animal, färsk förorening: HMBif, PLBif, TLBif, Pig2Bac, HF183TaqMan (100%) Fyra föroreningskällor, färsk förorening: BacR Pig2Bac AllBac FEqPCR CWMit PLMit (99.04%) Human vs. Animal äldre, utspädd förorening: NoV; PGMit (88.49%) Fyra föroreningskällor, äldre, utspädd förorening: NoV, TLBif/HMBif, TLBif/CWBif, TLBif/PLBif (82.78%)

8 Verktygslåda för fekal källspårning på laboratoriet och i fält, kontakt: Rikard Dryselius, Livsmedelsverket Fekal förorening Ingen fekal förorening Kommunalt avlopp Enskilt avlopp Humanfekalier Orenat avlopp Slam Djurfekalier Tamdjur Dagvatten Vilda djur Nöt Får Svin Hund Kyckling Häst Fåglar Vildsvin Betesmark eller gödsel Gäss Duvor Måsar

9 Metoder

10 Kemiska markörer, badvatten Nybro 8,00 7,00 6,00 E. coli Enterokocker Clostridiesporer Kolifager F-specifika fager Log halt/100 ml 5,00 4,00 3,00 2,00 1,00 0,

11 Mikrobiologiska markörer Analys från fekaliebibliotek (ca. 300 fecesprover från olika djurslag).

12 Sekvensering (16S amplikonsekvensering)

13 Source proportion Source Calf Cow Dog Poultry Horse Pig Sewage Sheep Storm water Wild bird Unknown 6. Broken sewage pipe 9. Heavy rain Events of contamination

14 Preliminär utvärdering verktygslådan From the results obtained this far, LC-MS/MS and metagenomics seem to be the most powerful approaches for efficient determination of faecal pollution (Dryselius et al. 2015)!

15 Utmaningar Koncentreringsmetoder, e.g. av virus och bakteriofager, sekundär konc. Limit of quantification (qpcr), i.e. volym analyserad med molekylära metoder Inbördes utbyte/detektionsnivå, vid användandet av fler markörer Enskilda avlopp! Ingen metod standard på vattenlab.