Nonylphenol Degradation During Wastewater Treatment -

Importance of Nitrifying Bacteria

Nonylphenol Degradation During Wastewater Treatment -

Importance of Nitrifying Bacteria

Presented By: Natasha SinhaPresented By: Natasha Sinha

Motivation: USGS Surface Water Reconnaissance(Kolpin et al., 2002)

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 Secondary Effluent Chromotograph

Further Motivation:Intersex Characteristics in Fish PopulationsFurther Motivation:Intersex Characteristics in Fish Populations

These data represent the frequency of intersex characteristics of fish taken above and below wastewater treatment discharges

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

Known Estrogenic Compounds in Wastewater

Conventional Wastewater Treatment

Processes Conventional Wastewater Treatment

Processes

Removal of Estrogens in Arizona Plants 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 10-10 < 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 BacteriaBackground 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.

• 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 SetupLab Setup

Reactor Synthetic Wastewater

Clarified Effluent

Control Unit

Variable: Sludge Age

Experimental Procedure Part 1Experimental Procedure Part 1

• Step 1: Look for NH4+ stimulation of O2 use

– Determine whether ammonia oxidizing bacteria are present.

• Step 1: Look for NH4+ stimulation of O2 use

– Determine whether ammonia oxidizing bacteria are present.

Result (1)Result (1)

t(s) vs. % Oxygen: Cells + Ammonia

0

10

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60

0 50 100 150 200 250 300

t(s)

% Oxygen

Cells + Ammonia 2

Cells + Ammonia 3

t(s) vs % Oxygen: Cells

0

10

20

30

40

50

60

70

0 50 100 150 200 250 300 350

t(s)

% Oxygen

t(s) vs. % Oxygen: Cells + Glucose

0

10

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80

0 50 100 150 200 250 300

t(s)

% Oxygen

Cells + Glucose 1

Cells + Glucose 2

OUR: 0.00059

OUR: Oxygen Utilization Rate

OUR: 0.00054

OUR: 0.00025

Measurements of nonylphenol degradation were taken after 24 hours and 48 hours using a C-18 procedure and Gas Chromatography

Was Nonylphenol Degraded?Was Nonylphenol Degraded?

No NP + NP

No ATU

+ ATU

Removal of Nonylphenol + DataRemoval of Nonylphenol + Data

To ComeTo Come

• Repeat with longer sludge retention time

• Compare to previous results

• Supplement with effluent NH4+ data

• Repeat with longer sludge retention time

• Compare to previous results

• Supplement with effluent NH4+ data

Acknowledgements Acknowledgements

Dr. Robert Arnold

Dave Newman

Bingfeng Dong

Dustin Brown & Arthur Ho

Dr. Robert Arnold

Dave Newman

Bingfeng Dong

Dustin Brown & Arthur Ho