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Tomato plant roots/ carboxymethyl cellulose (CMC) method for the removal and recovery of phosphate from agricultural wastewater University of Windsor Chemistry and Biochemistry David Ure 1 , Angela Awada 1 , Nicole Frowley 1 , Neils Munk 2 , Amanda Stanger 2 and Bulent Mutus 1 1. University of Windsor 2. Bruce Peninsula Biosphere Association

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Page 1: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Tomato plant roots/ carboxymethyl cellulose (CMC) method for the removal

and recovery of phosphate from agricultural wastewater

University of Windsor

Chemistry and Biochemistry

David Ure1, Angela Awada1, Nicole Frowley1, Neils Munk2, Amanda Stanger2 and Bulent

Mutus1

1. University of Windsor 2. Bruce Peninsula Biosphere Association

Page 2: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

The goal of this project: Reduce nutrient loadings from agricultural streams before it drains into rivers and lakes. Accumulation in aquatic ecosystems leads to eutrophication.

Page 3: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Eutrophication Eutrophication occurs when ecosystems accumulate too many nutrients

Aids the formation of harmful algal blooms algal toxins> dead algae> bacteria> depletion of oxygen> kill of aquatic animals

Page 4: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Causes many, our focus = P Phosphate is important in many aspects of biology Therefore, it is used in fertilizers

Like crops, algae depends on phosphate for growth and development Therefore, the accumulation of phosphate promotes algal growth

Page 5: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Phosphate Binders Chitosan matrix with chelated iron

From shell fish exoskeleton

Page 6: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Phosphate Binders Chitosan matrix with chelated iron

Drastic price increase Leeching of iron to environment Unstable: digested by microorganisms

Page 7: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Phosphate Binders Ideal binding matrix should; 1. Bind large quantities of phosphate 2. Inexpensive 3. Safe for the environment

Tomato plant roots

Page 8: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Binding Capacity

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70P-

Rec

over

ed (g

/kg)

Page 9: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Phosphate Binders Ideal binding matrix should; 1. Bind large quantities of phosphate 2. Inexpensive 3. Safe for the environment

Tomato plant roots

Probably

Page 10: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Plant Roots – Phosphate Binding Capacity: ~ 55 g/kg

Page 11: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Plant Roots – Phosphate Binding Capacity: ~ 55 g/kg

P

Page 12: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Tightness of Binding

KD App

Iron-Chito: 138.3mM

Roots: 23.1mM

Page 13: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Methods of P Recovery

Direct reapplication of P-saturated roots as

fertilizer

Page 14: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Recovery and Regeneration of Binding Capacity

P replaced with counter anion – maybe liquid fertilizer P - P - Typically, acid/base or

conjugate salt

Page 15: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Carboxymethyl Cellulose (CMC)

Page 16: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

P

CMC

CMC displaces P

P can be collected and the root material reused

Page 17: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

CMC enhances calcium-phosphate precipitation

Less calcium to harvest P

Lower cost

Precipitation of P as Calcium-Phosphate

Page 18: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

No CMC

With CMC Increasing calcium

(0-10mM)

Page 19: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Lion’s Head On Site Testing

Page 20: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Lion’s Head On Site Testing

Water source on site contaminated with manure.

Page 21: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Cham

ber 1

Cham

ber 2

Cham

ber 3

Direction of flow -Radial filtration

Biofilter-Idea: Neils Munk; Design: Mutus; Build: UW Tech Support Centre

Page 22: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Flow rate: ~4L/min

ISCO samplers

inflow

outflow inflow outflow

Page 23: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Lion’s Head On Site Testing

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[P] (

µg/L

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Iron-Sawdust Phosphate Levels – Before and After Filtration

Before After

Page 24: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Lion’s Head On Site Testing

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Iron-Sawdust Nitrate Levels – Before and After Filtration

Before After

Page 25: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Summary

Phosphate Removal (%)

Nitrate Removal (%)

Iron-Sawdust 26.2% 58.5%

Page 26: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

0100200300400500600700800

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phat

e (µ

g/L)

Roots Phosphate Levels – Before and After Filtration

Before After

Lion’s Head On Site Testing

Page 27: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Lion’s Head On Site Testing

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Root Nitrate Levels – Before and After Filtration

Before After

Page 28: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Summary

Phosphate Removal (%)

Nitrate Removal (%)

Iron-Sawdust 26.2% 58.5%

Plant Roots 70.8% 58.2%

Page 29: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

What in plant roots binds phosphate?

In collaboration with Dr. Robert Schurko, Austin Peach and Michelle Quan

Used solid state nuclear magnetic

resonance

Probe the chemical environment of P

atom

Page 30: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

ssNMR of Plant Roots Throughout Binding-Elution Cycle

Demonstrates strong uptake and removal of phosphate throughout the binding-removal cycle

Page 31: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

ssNMR of Plant Roots Throughout Binding-Elution Cycle

Not definitive on what binds P Co-run Na and H atom NMR

Page 32: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Iron-chito – iron interacts with P

P (-) Fe (+)

The NMR signal of metal – P interactions is distinct and not occurring in roots

NMR data suggests the formation of H-bond(s)

Page 33: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

H-bonds suggest binding to a protein All organisms, including tomatoes have phosphate transporter proteins that internalize phosphate

Outside

P

P P

P

P P

Inside

Page 34: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

P

P P

P

P

P P

Outside Inside

Page 35: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

P

P P

P

P

P P

Outside Inside

Page 36: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

P

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P

P

P P

Outside Inside

Page 37: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

P

P

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Page 38: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

P

P

P

P

P P

Outside Inside

Our roots are dead

No energy available

P

But, the P binding protein works without

energy

Page 39: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

P

Phosphate binds

Page 40: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Phosphate binds

Induce release with counter anions (CMC

or Cl)

P

Page 41: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Phosphate binds

Induce release with counter anions (CMC

or Cl)

Repeat

P - P -

Page 42: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

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Untreated Urea

PBC

Disrupt proteins with urea or heat treatment

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Untreated 2hr HeatPB

C

6 M Urea 150 °C

Page 43: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Current and Future Objectives

Phosphate transporter protein expression is regulated by phosphate in soil Study the effect on phosphate treatment on later use of roots for remediation Working with Holy Names High School – STEM Academy (Ms Mary-Ellen Kavanaugh)

Page 44: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Current and Future Objectives

P and No P roots

Flow rates

Page 45: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

0102030405060708090

P-R

ecov

ered

(g/k

g)

Different Ages and Types of Plant Roots

Page 46: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Current and Future Objectives

Study different plant roots ie. cucumber Study the degradation in binding capacity overtime

Page 47: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Acknowledgements: WECO2 Parks: Matthew Posthumus, Sergey Postnikov Bruce Peninsula Biosphere Association (BPBA): Katherine McLeod, Amanda Stanger, Elizabeth Thorn and Neils Munk Essex Region Conservation Authority (ERCA): William Tate, Samantha Dundas, Mackenzie Porter, Katie Stammler and Kevin Money Funding: OMAFRA, BPBA, NSERC-ENGAGE, MITACS, UTRCA, ECCC, University of Windsor Alumni Association (Anonymous Donor)

Page 48: University of Windsor - Latornell Conservation Symposium · University of Windsor 2. Bruce Peninsula Biosphere Association . The goal of this project: Reduce nutrient loadings from

Thank you very much for listening Questions? Bulent Mutus: [email protected] David Ure: [email protected]