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By: Alyssa Ruiz

Mentor: Dr. Tesfa Yacob

PI: Dr. Karl Linden

REINVENT THE TOILET CHALLENGE: URINE TREATMENT

ENVIRONMENTAL SUSTAINABILITY REU 2013

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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

ABOUT THE CHALLENGE

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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

Urea() Total Ammonia majority of total N compounds causes pH increase more = higher inactivation

BACKGROUND: URINE

hydrolysis

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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 = Fertilizer

ABOUT THE SYSTEM

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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 ammonia2. the intensity of heat treatment3. the duration of the application of heat treatment

Assess the potential for regrowth in urine

0BJECTIVES

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Two surrogates used from frozen cultures E. coli Famp MS2

Enumeration Methods Membrane Filtration - E. coli Famp Agar Plating - MS2

MATERIALS AND METHODS

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Initial Experiment: 40°C & 60°C over 1 hr Surrogates: E. coli & MS2Total Ammonia Concentration: 2 g/L

EXPERIMENTS: PRELIMINARY DISINFECTION

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RESULTS: PRELIMINARY DISINFECTION

0 10 20 30 40 50 60 700

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2

3

4

5

6

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Urine spiked with E. Coli Famp, Heat Disinfection, 2 g/L TNH3

40 °C

60 °C

Time (minutes)

log

CFU/

10

mL

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RESULTS: PRELIMINARY DISINFECTION

0 1 0 2 0 3 0 4 0 5 0 6 0 7 00

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2

3

4

5

6

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Urine Spiked with MS2, heat Disinfection at 60°C, 2g/l tNH3

Time (minutes)

Log

pFU/

10 m

L

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Previous experiment shows 60°C more than enough while literature says must be above 45°C

Maximize efficiency Chose 50°C because previous data implied should

take less than one day - optimalCurious about a synergistic effect

3 treatment methods AH: Ammonia + Heat H: Heat Only A: Ammonia Only

Tested over 24 hours2.3 g/L NH3Two surrogates used: E. coli Famp & MS2

EXPERIMENTS: 50°C DISINFECTION

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RESULTS: 50°C DISINFECTION

0 2 4 6 8 1 0 1 2 1 4 1 6 1 8 2 0 2 2 2 4 2 60

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2

3

4

5

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urine spiked with E. coli famp, heat Disinfection 50°C

AH

H

A

Time (hours)

log

CFU/

mL

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RESULTS: 50°C DISINFECTION

0 2 4 6 8 1 0 1 2 1 4 1 6 1 8 2 0 2 2 2 40

1

2

3

4

5

6

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urine spiked with ms2, heat Disinfection 50°C

AH

H

A

Time (hours)

log

pFU'

s/mL

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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 days10^4 starting E. coli concentration

EXPERIMENTS: REGROWTH

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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 0 01:1 3.99 ± 0.11 0 01:5 3.98 ± 0.15 0 0

TSB (pos) 4.38 ± 0.03 9.29 ± 0.13 NAPBS (neg) 4.07 ± 0.14 3.05 ± 1.71 2.56 ± 0.03

RESULTS: REGROWTH

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First of these types of experimentsSynergistic effect proven

significant in short term urine disinfection implications for other avenues: passive solar heating

Further test synergistic effect with varying ammonia concentrations

Regrowth experiments with spiking before heatShort, low heat + storageOther surrogates – Enterococcus, ascaris, eggs

CONCLUSIONS & FURTHER EXPERIMENTS

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NSF REU ProgramCU Boulder – Dr. MontoyaBill & Melinda Gates FoundationDr. Karl LindenDr. Tesfa YacobRyan MahoneySara BeckCori OversbyLinden/Gates Team

ACKNOWLEDGEMENTS

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"B i l l & Mel inda Gates Foundat ion."   Water, San i tat ion & Hyg iene . N .p . , n .d . Web. 8 June 2013.

<http : / /www.gates foundat ion.org/What -We-Do/G lobal -Deve lopment/Water-San i tat ion-and-Hygiene>.

"Hach – Manufactures Water Qual i ty Tes t ing and Analyt ica l Ins truments & Reagents ."  Hach –

Manufactures Water Qual i ty Test ing and Analyt ica l Ins t ruments & Reagents . N .p . , n .d . Web. 8 June 2013. <http : / /www.hach.com/>.

Höglund , C . e t a l . "Evaluat ion of Faecal Contaminat ion and Microb ia l Die -Off in Ur ine Separat ing Sewage Systems."   Water Sc ience & Technology  38 .6 (1998) : 17-25. Pr int . Putnam, David F. “Compos i t ion and Concentrat ive Propert ies of Human Ur ine .”

(1971) : n . pag.  Goog le Scholar . Web. McCartney, Dary l , and Kr is t ine M Wichuk. “A Review of the Effect iveness of Current

Time– Temperature Regulat ions on Pathogen Inact ivat ion Dur ing Compost ing .”   J ournal o f

Env i ronmental Eng ineer ing and Sc ience 6.5 (2007): 573–586.   CrossRef . Web. Udert , K . M. et a l . “Fate of Major Compounds in Source-separated Ur ine .”   Water

Sc ience & Technology  54.11–12 (2006) : 413-20. Pr int . Vinnerås , B jörn e t a l . “ Inact ivat ion of Bacter ia and Vi ruses in Human Ur ine

Depend ing on Temperature and D i lut ion Rate .”   Water Research  42 .15 (2008) : 4067–4074.   CrossRef .

Web.

REFERENCES

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“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

QUESTIONS?