Post on 06-Jun-2020
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
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.
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
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
Phosphate Binders Chitosan matrix with chelated iron
From shell fish exoskeleton
Phosphate Binders Chitosan matrix with chelated iron
Drastic price increase Leeching of iron to environment Unstable: digested by microorganisms
Phosphate Binders Ideal binding matrix should; 1. Bind large quantities of phosphate 2. Inexpensive 3. Safe for the environment
Tomato plant roots
Binding Capacity
0
10
20
30
40
50
60
70P-
Rec
over
ed (g
/kg)
Phosphate Binders Ideal binding matrix should; 1. Bind large quantities of phosphate 2. Inexpensive 3. Safe for the environment
Tomato plant roots
Probably
Plant Roots – Phosphate Binding Capacity: ~ 55 g/kg
Plant Roots – Phosphate Binding Capacity: ~ 55 g/kg
P
Tightness of Binding
KD App
Iron-Chito: 138.3mM
Roots: 23.1mM
Methods of P Recovery
Direct reapplication of P-saturated roots as
fertilizer
Recovery and Regeneration of Binding Capacity
P replaced with counter anion – maybe liquid fertilizer P - P - Typically, acid/base or
conjugate salt
Carboxymethyl Cellulose (CMC)
P
CMC
CMC displaces P
P can be collected and the root material reused
CMC enhances calcium-phosphate precipitation
Less calcium to harvest P
Lower cost
Precipitation of P as Calcium-Phosphate
No CMC
With CMC Increasing calcium
(0-10mM)
Lion’s Head On Site Testing
Lion’s Head On Site Testing
Water source on site contaminated with manure.
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
Flow rate: ~4L/min
ISCO samplers
inflow
outflow inflow outflow
Lion’s Head On Site Testing
0
50
100
150
200
250
300
350D
ay 1
Day
2D
ay 3
Day
4D
ay 5
Day
6D
ay 7
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ay 9
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[P] (
µg/L
)
Iron-Sawdust Phosphate Levels – Before and After Filtration
Before After
Lion’s Head On Site Testing
0500
10001500200025003000350040004500
Day
1D
ay 2
Day
3D
ay 4
Day
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ay 6
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ay 8
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ay 1
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ay 1
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ay 1
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ay 2
0D
ay 2
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ay 2
2D
ay 2
3D
ay 2
4
Nitr
ates
(µg/
L)
Iron-Sawdust Nitrate Levels – Before and After Filtration
Before After
Summary
Phosphate Removal (%)
Nitrate Removal (%)
Iron-Sawdust 26.2% 58.5%
0100200300400500600700800
Day
1D
ay 2
Day
3D
ay 4
Day
5D
ay 6
Day
7D
ay 8
Day
9D
ay 1
0D
ay 1
1D
ay 1
2D
ay 1
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ay 1
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ay 1
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ay 1
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ay 1
7D
ay 1
8D
ay 1
9D
ay 2
0D
ay 2
1D
ay 2
2D
ay 2
3
Phos
phat
e (µ
g/L)
Roots Phosphate Levels – Before and After Filtration
Before After
Lion’s Head On Site Testing
Lion’s Head On Site Testing
0
500
1000
1500
2000
2500D
ay 1
Day
2D
ay 3
Day
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ay 5
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ay 7
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ay 9
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24
Nitr
ate
(µg/
L)
Root Nitrate Levels – Before and After Filtration
Before After
Summary
Phosphate Removal (%)
Nitrate Removal (%)
Iron-Sawdust 26.2% 58.5%
Plant Roots 70.8% 58.2%
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
ssNMR of Plant Roots Throughout Binding-Elution Cycle
Demonstrates strong uptake and removal of phosphate throughout the binding-removal cycle
ssNMR of Plant Roots Throughout Binding-Elution Cycle
Not definitive on what binds P Co-run Na and H atom NMR
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)
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
P
P P
P
P
P P
Outside Inside
P
P P
P
P
P P
Outside Inside
P
P P
P
P
P P
Outside Inside
P
P
P
P
P P
Outside Inside
P
P
P
P
P
P P
Outside Inside
Our roots are dead
No energy available
P
But, the P binding protein works without
energy
P
Phosphate binds
Phosphate binds
Induce release with counter anions (CMC
or Cl)
P
Phosphate binds
Induce release with counter anions (CMC
or Cl)
Repeat
P - P -
0
10
20
30
40
50
60
70
Untreated Urea
PBC
Disrupt proteins with urea or heat treatment
0
10
20
30
40
50
60
70
Untreated 2hr HeatPB
C
6 M Urea 150 °C
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)
Current and Future Objectives
P and No P roots
Flow rates
0102030405060708090
P-R
ecov
ered
(g/k
g)
Different Ages and Types of Plant Roots
Current and Future Objectives
Study different plant roots ie. cucumber Study the degradation in binding capacity overtime
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)
Thank you very much for listening Questions? Bulent Mutus: mutusb@uwindsor.ca David Ure: ured@uwindsor.ca