Presented By: Natasha Sinha Nonylphenol Degradation During Wastewater Treatment - Importance of Nitrifying Bacteria Presented By: Natasha Sinha

Motivation:  USGS Surface Water Reconnaissance (Kolpin et al., 2002) Comparison of 4 Compounds: Compound (Use and Effect) CMAX (µg/L) Days to Med Dose Caffeine (stimulant) 5.7 5.7 x 103 (Excedrin Migraine) Ibuprofen (anti-inflammatory) 1.0 105 (Motrin) 17α-ethinyl estradiol (oral contraceptive) 0.27 37 (Alesse-21) 17β-estradiol (vertebrate hormone) 0.093 (Phoenix) 5.3 x 103 (Activella) 4-nonylphenol (surfactant metabolite) 40 (Tucson) 4.6 x 10-4 as estrogenic E2, EE2 Sampling regions for the USGS Reconnaissance of US surface water

Secondary Effluent Chromotograph

Further Motivation: Intersex Characteristics in Fish Populations These data represent the frequency of intersex characteristics of fish taken above and below wastewater treatment discharges

Relative Estrogenic Activity Known Estrogenic Compounds in Wastewater Chemical Structure Molecular Weight Log Kow Relative Estrogenic Activity (YES bioassay) 17-estradiol (E2) 272.39 3.94(9) 1.0 17-ethinyl estradiol (EE2) 296.40 4.15(9) 1.32 Estrone (E1) 270.37 3.43(9) 0.20 Estriol (E3) 288.39 2.81(9) 0.01 Nonylphenol (NP) 220.35 4.48(10) 2×10-4 Octylphenol (OP) 206.33 4.12(10) 3×10-4 Bisphenol A (BPA) 228.29 3.40(11) 8 x 10-5

Conventional Wastewater Treatment Processes

Removal of Estrogens in Arizona Plants Total Estrogenic Activity (M as EE2) Facility Influent Effluent Fraction Removal Avra Valley 1 x 10-9 2 x 10-12 0.998 Roger Road 3 x 10-10 2 x 10-10 0.33 Ina Road 4 x 10-11 5 x 10-12 0.88 Randolph Park < 2 x 10-12 > 0.99 Wildcat 1 x 10-10 4 x 10-12 0.96 Rio de Flag 1.5 x 10-10 < 6 x 10-13 > 0.996

Background on Ammonia Oxidizing Bacteria NH4+ NH2OH NO2 O2 AMO (ammonia monoxygenase) AMO is a non-selective monoxygenase Central Hypothesis: Estrogen removal is due to AMO activity in culture.

Lab Setup Synthetic Wastewater Control Unit Clarified Effluent Reactor Variable: Sludge Age

Experimental Procedure Part 1 Step 1: Look for NH4+ stimulation of O2 use Determine whether ammonia oxidizing bacteria are present.

Result (1) OUR: Oxygen Utilization Rate OUR: 0.00054 OUR: 0.00059

Was Nonylphenol Degraded? Measurements of nonylphenol degradation were taken after 24 hours and 48 hours using a C-18 procedure and Gas Chromatography No NP + NP No ATU + ATU

Removal of Nonylphenol + Data

To Come Repeat with longer sludge retention time Compare to previous results Supplement with effluent NH4+ data

Dustin Brown & Arthur Ho Acknowledgements Dr. Robert Arnold Dave Newman Bingfeng Dong Dustin Brown & Arthur Ho