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Fate of Chemical and Biological Emerging Contaminants in Biosolids and After Land Application

Fate of Chemical and Biological Emerging Contaminants in Biosolids and After Land Application. Ian L. Pepper The University of Arizona. The Northwest Biosolids Management Association’s 23 rd Annual Biosolids Management Conference September 19-21, 2010 Campbell’s Conference Center Chelan, WA.

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Fate of Chemical and Biological Emerging Contaminants in Biosolids and After Land Application

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  1. Fate of Chemical and Biological Emerging Contaminants in Biosolids and After Land Application Ian L. PepperThe University of Arizona The Northwest Biosolids Management Association’s 23rd AnnualBiosolids Management ConferenceSeptember 19-21, 2010Campbell’s Conference CenterChelan, WA

  2. EMERGING CONTAMINANTS ● Include chemical and biological entities ● By definition are newly discovered contaminants ● Little information on incidence, fate and transport, and potential adverse impacts on human and/or ecological health

  3. CHEMICAL EMERGING CONTAMINANTS Endocrine Disrupting Compounds (EDCs) ● Pharmaceuticals and personal care products (PPLP) ● Steroidal hormones (estrogenic activity)● Flame retardants (polybrominated diphenyl ethers (PBDEs)

  4. Representative Estrogenic Compounds

  5. Polybrominated Diphenyl ethers Brm Brn . . . . . . . . . . . . . . • Flame retardants • 229 possible congeners

  6. Characteristics of EDCs. ● EDCs interfere with the synthesis, secretion, transport, binding, action, or elimination of natural hormones in the body that are responsible for the maintenance of homeostasis (normal cell metabolism), reproduction, development, and/or behavior. ● EDCs can be hormone mimics, with hormone-like structures and activities. That is EDCs sometimes have chemical properties similar to hormones and bind to hormone specific receptors in or on the cells of target organs. ● EDCs frequently have lower potency than the hormones they mimic (i.e., require higher dose to elicit an equivalent response), but may be present in water at high concentrations relative to natural hormones. Furthermore, EDCs may not be subject to normal (internal) regulations mechanisms. ● For all known EDCs, there is some dose below which there is no observable response.

  7. Most potential adverse effects of EDCs focus on wildlife or ecosystems. ● Eggshell thinning and subsequent reproductive failure of waterfowl ● Reduced populations of Baltic seals ● Reproductive failure in alligators ● Development of male sex organs in female marine animals such as whelks and snails ● Reduced or malformed frog populations ● Disruption of normal sex ratios among exposed populations of fish

  8. THE LANDMARK USGS 1999-2000 STUDY ● Barnes et al. (2002) water-quality data for phamaceuticals, hormones, and other organic wastewater contaminants in U.S. streams, 1999-2000. USGS Open-File Report

  9. Locations of stream sampling sites in the 1999–2000 reconnaissance of United States surface water quality by USGS.

  10. Hormones and hormone mimics observed in U.S. surface waters.

  11. Representative pharmaceuticals measured in the 1999–2000 UGS reconnaissance of U.S. streams. A comparison of drinking water levels with medicinal doses. aThe mass of caffeine in two Excedrin tablets. There is 135 mg of caffeine in an 8-oz. cup of coffee.bThe mass of ibuprofen in two tablets of AdvilcThe lowest adult daily dose of cimetidinedRange of 17 -ethinyl estradiol masses in birth control pillseRange of testosterone masses provided over 3–6 months when used for hormone replacementfRecommended adult dosages

  12. Physical, chemical and biochemical properties of selected estrogenic chemicals.

  13. SUSTAINABILITY OF LAND APPLICATION Huruy Zerzghi University of Arizona

  14. STUDY SITE • Study begun in 1986 at the U of A (Marana Agricultural Center) • Biosolids has been continually applied for 20 years (1986 -2005) • Crop grown - cotton Marana

  15. DESIGN OF THE EXPERIMENT • Control (# 1) • Inorganic fertilizer (# 3) • Low biosolids rate (1x, # 2) • High biosolids rate (3x, # 4)

  16. The twentieth year of biosolids land application (March 1-15, 2005) 8% solids

  17. Biosolids land application

  18. FATE OF LAND APPLIED ENDOCRINE DISRUPTORSIN SOIL

  19. Mean congener (BDE-47, -99, and -209) concentrations in soil in the 0X, 1X, and 3X biosolid loading rate plots.

  20. Total mean concentration of PBDE congeners* in soils at the 0-30 cm soil depth compared to household indoor exposure. From Fromme et al., 2009.*For BDE-47, -99, -153 and -209

  21. Nonylphenol Soil depth 0-1’

  22. RISK ASSESSMENT FOR EXPOSURE TO PBDEChuck Gerba Dermal Adsorption Daily Exposure Dose Dermal Intake = [(C)(BSA)(SAS)(AF)(OEF)] / [(BS)(1,000)] Where BSA = body surface area (cm2/day), SAS = soil adhered to the skin (mg/cm2), AF = fraction of PBDE adsorbed to the skin, and BW = body weight. Total exposure is the sum of ingestion and dermal intake.

  23. RISK ASSESSMENT FOR EXPOSURE TO PBDEChuck Gerba Ingestion Daily Exposure Dose Daily ingestion exposure dose = [(C) (SIR) (OEF)] /BW Where C =PBDE concentration (ng/g dry weight), SIR = soil ingestion rate (m3/day), BW = body weight (kg), and OEF = outdoor exposure fraction (hours spent over a day in the field to which biosolids have been added).

  24. Evaluation of hazard indices for BDE 47, 99, 153 and 209. Hazard index is summation of Hazard Quotient values.

  25. RISK Non Cancer Risk ● Hazard Index is several orders of magnitude < 1 suggesting very low risk Cancer Risk ● BDE 209 is only congener where information available to evaluate carcinogenicity - Mean concentration for 3X plot = 61.4 ng/g - Dermal and ingestion exposure = 0.0197 ng/kg/day6Life Time Risk of Cancer = 9.11 x 10-14

  26. SURVIVAL OF PRIONS IN CLASS B BIOSOLIDSKazue Takizawa, Chuck Gerba, and Ian Pepper The University of Arizona

  27. Kazue Takizawa ● M.S. degree in SWES● Optimized extraction of prion proteins from Class B biosolids● Evaluated survival of prions in Class B biosolids at mesophilic and thermophilic temperatures

  28. Prion Characteristics ● A normal prion (PrPc) is composed of mostly alpha-helicies ● An infectious prion (PrPsc) is dominated by beta-sheets ● Resistant to inactivation by proteolytic enzymes, conventional disinfectants, and standard sterilization methods http://www.le.ac.uk/biology/research/phyto/prions.jpg

  29. Infectious Prion Conversion PrPc PrPsc ● When a infectious prion PrPsc comes in contact with a normal prion PrPc, it causes PrPc to convert to PrPsc ● Disease occurs when PrPsc concentration threshold is reached

  30. Prion Pathogenesis ● Infectious prion proteins (PrPsc) cause Transmissible Spongiform Encephalopathy (TSE) diseases which affects humans and animals ● Most common Creutzfeldt-Jakob disease (CJD) Soto et al. 2004

  31. TSEs in Animals ●Affects sheep, goats, minks, mules, deer, cows, cats, exotic felines, and ungulates ● Most common TSE in animals is called scrapie which affects sheep and goats ● Chronic Wasting Disease (CWD)- affecting deer and elk - Affecting deer population in U.S. :Colorado, Wyoming, Nebraska, New Mexico, South Dakota, Wisconsin

  32. FATE OF PRIONS IN THE ENVIRONMENT ● PrPsc adsorbs to clay and organic colloids● Sorbed prions reported to remain infective● Prions reported to survive mesophilic anaerobic digestion during wastewater treatment● Detection via Western Blot technology—did not assess infectivity

  33. POTENTIAL ROUTE OF EXPOSURE TO PRIONS VIA LAND APPLICATION Animal Slaughterhouse Wastewater Wastewater Treatment Biosolids Humans Cows Land Application

  34. METHODOLOGY ● Class B biosolids—7% solids● CAD 1A2DS (CAD5) cell line● Source of prions: Rocky Mountain Laboratory (RML)—infected mouse brain● Enzyme-linked immunosorbent spot (ELISPOT) assay

  35. Approximately 1,000 prion infected CAD5 cells The original plan was to use ScN2a cells which yielded 0.001% infectivity rate ● Currently, ELISPOT assays performed with CAD5 cells has dramatically increased the yield to a 50% infectivity rate. ● Decreased assay by 8 days

  36. EVALUATION OF EXTRACTANTS OF PRION PROTEINS FROM BIOSOLIDS ● Phosphate buffer saline (PBS)● 3% beef extract● Sodium dodecyl sulfate (SDS)● 8M urea

  37. RECOVERY OF PRIONS FROM BIOSOLIDS

  38. REDUCTION OF INFECTIOUS PRIONS AT MESOPHILIC (37◦C) AND THERMOPHILIC (60◦C) TEMPERATURES A) In PBS Mesophilic: log10 N/N0 (15 days) = ‐1.13 Thermophilic: log10 N/N0 (10 days) = ‐1.80B) Class B biosolids Mesophilic: log10 N/N0 (15 days) = ‐2.43 Thermophilic: log10 N/N0 (10 days) = ‐3.41

  39. INACTIVATION OF PRIONS IN BIOSOLIDS > PBS ● Mechanism: protolytic enzymes that denature proteins● Ammonia or other substances in biosolids

  40. COMPARISON OF STUDIES ● Maluquer de Motes et al. (2008) - prion survival in raw sewage - Western blot technology - 90% reduction of prions after 32.6 days at 20◦C● Our study - 99.1% reduction after 15 days at 37 ◦C - 99.9% reduction after 10 days at 60 ◦C Reason: infectivity assay

  41. SUMMARY ● 4M urea an effective extractant of prions from biosolids● Significant inactivation in biosolids at mesophilic and thermophilic temperatures ● Data needed on inactivation during actual wastewater treatment

  42. ONGOING PRION WORK ● Evaluating wastewater treatment effects on prion inactivation - miniature anaerobic digesters (test tubes) - mesophilic anaerobic digestion 37°C - thermophilic anaerobic digestion 60°C - composting

  43. FATE OF EMERGING CONTAMINANTS: CONCLUSIONS ● Estrogenic compounds added to soil via land application of biosolids degrade quickly within a matter of weeks ● PBDEs are strongly hydrophobic and sorbed to soil colloids with low solubility and bioavailability ● Overall risk from PBDE via land application is very low. ● Prions are inactivated in biosolids at mesophilic and thermophilic temperatures ● Overall more research is needed in both areas of concern

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