1. © NERC All rights reserved Organic Sewage Tracers Sterols are components of cells & are synthesized by most organisms In humans cholesterol is biohydrogenated by intestinal microflora to 5  cholestrol Other animals also produce 5  cholestrol but at much lower concentrations Herbiverous animals produce 24-ethyl coprostanol in greater concentrations Sterol structure

2. © NERC All rights reserved Assessment of Faecal Contamination Using Sterol Molecular Markers Attenborough Ponds Nature: Long Term Effects of River Diversion

3. © NERC All rights reserved Conneries dating 1986 ? 1963 ?

4. © NERC All rights reserved TOC vs Sterol Concentration

5. © NERC All rights reserved Identification of Sewage Markers Target mix TIC m/z = 333 Cholesterol-d6 m/z = 231 Coprostan-3-one m/z = 329 Cholesterol Resolving co-elution Sediment Sample coprostanol m/z 333 m/z 231 m/z 329 estrone  estradiol 5  - cholestane coprostanol 5  -epicoprostanol 5  - cholestanol campesterol stigmasterol fucosterol +  sitosterol 5  -stigmastanol Environment & Human Health Theme

6. © NERC All rights reserved Ten of eleven sewage marker compounds were identified in the Attenborough cores 5  -cholestane not detected

7. © NERC All rights reserved Sterol & Stanol Marker Pathways Intestinal (5  ) Environmental (5  )

8. © NERC All rights reserved 1. Sitosterol: Clifton and Church = Input from plants & or avian FM 2. Stigmastanol: Conneries = Input from ruminants / manure, slurry ? 3. Coprostanol: Concs. >0.5ug/g = received appreciable amounts of sewage 1 2 3

9. © NERC All rights reserved

11. 1. Coprostanol/Cholesterol: human sewage >0.2 criteria= Only Conneries 2. Coprostanol/(Coprostanol+Cholestanol) human sewage >0.7= Conneries 3. Epicoprostanol/coprostanol:treatment or age in environ. =Church & Clifton little sewage or high degree of treatment

12. © NERC All rights reserved 1.5 1.0 0.5 0.0 0.51.01.52.02.50.0 Low sewage, old or treated High sewage, untreated Bi-plot of sewage marker compounds

13. © NERC All rights reserved

14. Ratio 5  Cholestanol/Cholesterol:anaerobic process marker Conneries possible more reducing environ at 20 to 50 cm ?