Final Research Presentation - University of Colorado Boulder

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REINVENT THE
TOILET CHALLENGE:
URINE TREATMENT
ENVIRONMENTAL
SUSTAINABILITY REU
2013
By: Alyssa Ruiz
Mentor: Dr.
Tesfa Yacob
PI: Dr. Karl
Linden
1
ABOUT THE CHALLENGE
 2.5 billion people practice open defecation or lack adequate
sanitation facilities
 Poor sanitation contributes to 1 .5 million child deaths from diarrhea
each year
 Main Objectives
 radically change our world’s current sanitation state through
innovative discoveries that convert our waste into viable resources
 decrease the statistic of child mortality that societies around the globe
are facing and to improve the quality of life in these communities
2
BACKGROUND: URINE
 Contains the majority of the nutrients (N & P) excreted
from our system
 Approximately 158 different chemical components,
including electrolytes, nitrogenous compounds, vitamins,
hormones, organic acids, and other various compounds
 Focus on Ammonia
 present after microbial urea hydrolysis
hydrolysis
 Urea( 𝑁𝐻2 𝐶𝑂 𝑁𝐻2 )
Total Ammonia
 majority of total N compounds
 causes pH increase
 more = higher inactivation
3
ABOUT THE SYSTEM
 Source separated toilet
 utilize the properties found in
urine
 create biochar
 energy efficient
 Main source of pathogens in
source separated urine comes
from misplaced feces
 Source separated urine high
potential for regrowth
 Less water compared to
latrines
 Daily sanitation
 End Product = Fer tilizer
4
0BJECTIVES
 Experiments have shown disinfection over long periods of time
(weeks) at low temperature and varying ammonia
concentrations
 Assess short term (hours) disinfection with these variables
1. the optimum concentration of ammonia
2. the intensity of heat treatment
3. the duration of the application of heat treatment
 Assess the potential for regrowth in urine
5
MATERIALS AND METHODS
 Two surrogates used from frozen cultures
 E. coli Famp
 MS2
 Enumeration Methods
 Membrane Filtration - E. coli Famp
 Agar Plating - MS2
6
EXPERIMENTS: PRELIMINARY
DISINFECTION
 Initial Experiment: 40 °C & 60°C over 1 hr
 Surrogates: E. coli & MS2
 Total Ammonia Concentration: 2 g/L
7
RESULTS: PRELIMINARY DISINFECTION
URINE SPIKED WITH E. COLI FAMP, HEAT DISINFECTION,
2 g/L TNH3
10
9
LOG CFU/ 10 mL
8
7
40 °C
6
5
4
60 °C
3
2
1
0
0
10
20
30
40
50
60
70
TIME (MINUTES)
8
RESULTS: PRELIMINARY DISINFECTION
URINE SPIKED WITH MS2, HEAT DISINFECTION AT 60 °C, 2g/L TNH3
8
7
LOG PFU/10 mL
6
5
4
3
2
1
0
0
10
20
30
40
50
60
70
TIME (MINUTES)
9
EXPERIMENTS: 50°C DISINFECTION
 Previous experiment shows 60 °C more than enough while
literature says must be above 45 °C
 Maximize ef ficiency
 Chose 50°C because previous data implied should take less
than one day - optimal
 Curious about a synergistic ef fect
 3 treatment methods
 AH: Ammonia + Heat
 H: Heat Only
 A: Ammonia Only
 Tested over 24 hours
 2.3 g/L NH3
 Two surrogates used: E. coli Famp & MS2
10
RESULTS: 50°C DISINFECTION
URINE SPIKED WITH E. COLI FAMP, HEAT DISINFECTION 50 °C
7
6
LOG CFU/mL
5
AH
4
3
H
2
A
1
0
0
2
4
6
8
10
12
14
16
18
20
22
24
26
TIME (HOURS)
11
RESULTS: 50°C DISINFECTION
URINE SPIKED WITH MS2, HEAT DISINFECTION 50 °C
7
6
LOG PFU'S/mL
5
AH
4
3
H
2
A
1
0
0
2
4
6
8
10
12
14
16
18
20
22
24
TIME (HOURS)
12
EXPERIMENTS: REGROWTH
 Factors that could support regrowth
 Contamination
 High abundance of nutrients found in urine
 Tested 3 urine dilutions
 1:0 (2.4 g/L NH3)
 1:1 (1.2 g/L NH3)
 1:5 (0.4 g/L NH3)
 Tested 2 controls
 Positive: TSB growth media
 Negative: PBS
 Designed to run for 21 days
 10^4 starting E. coli concentration
13
RESULTS: REGROWTH
Sample
Day 0 (avg log
CFU/ 1 mL)
Day 2 (avg log
CFU/ 1 mL)
Day 24 (avg log
CFU/ 1 mL)
1:0
3.95 ± 0.14
3.99 ± 0.11
3.98 ± 0.15
4.38 ± 0.03
4.07 ± 0.14
0
0
0
0
0
0
9.29 ± 0.13
NA
3.05 ± 1.71
2.56 ± 0.03
1:1
1:5
TSB (pos)
PBS (neg)
14
CONCLUSIONS & FURTHER EXPERIMENTS
 First of these types of experiments
 Synergistic ef fect proven
 significant in short term urine disinfection
 implications for other avenues: passive solar heating
 Further test synergistic ef fect with varying ammonia
concentrations
 Regrowth experiments with spiking before heat
 Short, low heat + storage
 Other surrogates – Enterococcus, ascaris, eggs
15
ACKNOWLEDGEMENTS









NSF REU Program
CU Boulder – Dr. Montoya
Bill & Melinda Gates Foundation
Dr. Karl Linden
Dr. Tesfa Yacob
Ryan Mahoney
Sara Beck
Cori Oversby
Linden/Gates Team
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REFERENCES
 " B i l l & M e l i n d a G a te s Fo u n d a t i o n . " Wa te r, S a n i t a t i o n & H yg i e n e . N . p . , n . d . We b . 8 J u n e
2 01 3 .
 < h t t p : / / w w w. g a t e s fo u n d a t i o n . o r g / W h a t - We - D o / G l o b a l - D e v e l o p m e n t / Wa t e r - S a n i t a t i o n and-Hygiene>.
 " H a c h – M a n u f a c t u r e s Wa te r Q u a l i t y Te s t i n g a n d A n a l y t i c a l I n s t r u m e n t s &
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 M a n u f a c t u r e s Wa te r Q u a l i t y Te s t i n g a n d A n a l y t i c a l I n s t r u m e n t s & R e a g e n t s . N . p . , n . d .
We b . 8 J u n e 2 01 3 . < h t t p : / / w w w. h a c h . c o m / > .
 H ö g l u n d , C . e t a l . " E v a l u a t i o n o f Fa e c a l C o n t a m i n a t i o n a n d M i c r o b i a l D i e - O f f i n U r i n e
 S e p a r a t i n g S e wa g e S y s te m s . " Wa te r S c i e n c e & Te c h n o l o g y 3 8 . 6 ( 1 9 9 8 ) : 17 - 2 5 . P r i n t .
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 M c C a r t n ey, D a r y l , a n d K r i s t i n e M W i c h u k . “A Re v i e w o f t h e E f fe c t i v e n e s s o f C u r r e n t
Time–
 Te m p e r a t u r e Re g u l a t i o n s o n P a t h o g e n I n a c t i v a t i o n D u r i n g C o m p o s t i n g . ” J o u r n a l o f
E n v i r o n m e n t a l E n g i n e e r i n g a n d S c i e n c e 6 . 5 ( 2 0 07 ) : 57 3 – 5 8 6 . C r o s s R e f . We b .
 U d e r t , K . M . e t a l . “ Fa te o f M a j o r C o m p o u n d s i n S o u r c e - s e p a r a t e d U r i n e . ” Wa te r S c i e n c e
&
 Te c h n o l o g y 5 4 . 1 1 – 1 2 ( 2 0 0 6 ) : 41 3 - 2 0 . P r i n t .
 V i n n e r å s , B j ö r n e t a l . “ I n a c t i v a t i o n o f B a c te r i a a n d V i r u s e s i n H u m a n U r i n e D e p e n d i n g o n
 Te m p e r a t u r e a n d D i l u t i o n R a te . ” Wa te r R e s e a r c h 4 2 . 1 5 ( 2 0 0 8 ) : 4 0 6 7 – 4 074 . C r o s s R e f .
We b .
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QUESTIONS?
“OUR GOAL: to enable universal access to sustainable
sanitation services by supporting the development of radically
new sanitation technologies as well as markets for new
sanitation products and services .” – Gates Foundation
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