![Page 1: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/1.jpg)
Stewardship and the Environment - Wastewater Treatment and Antibiotics
Timothy M. LaParaDepartment of Civil, Environmental,
and Geo- EngineeringUniversity of Minnesota
4 November 2015
![Page 2: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/2.jpg)
http://www.cdc.gov/drugresistance/threat-report-2013/
![Page 3: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/3.jpg)
![Page 4: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/4.jpg)
Antimicrobial resistance is a complex problem driven by many
interconnected factors; single, isolated interventions have little
impact.
Pruden et al. 2013 Environ. Health Persp. 121:878-885. WHO. 2012. www.who.int/mediacentre/factsheets/fs194/en/41
![Page 5: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/5.jpg)
How is antibiotic resistance detected/quantified?
Medical Approach:1. Isolate pathogen from a patient2. Determine which antibiotics are effective
Model Organism Approach:1. Isolate a model organism 2. Determine which antibiotics are effective
(Great Plate Count Anomaly?)
![Page 6: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/6.jpg)
New Methodology
• Target genes quantified by real-time polymerase chain reaction
Cycle Number
Fluo
rese
nce
105 g
enes
104 g
enes
103 g
enes
102 g
enes
6.6×104 genes
3.1×102 genes
![Page 7: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/7.jpg)
The “ARG” Cycle
http://www.cdc.gov/drugresistance/threat-report-2013/
![Page 8: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/8.jpg)
1019 copies/day
1017 copies/day
1018 copies/day
Auerbach et al. 2007. Water Research 41:1143–1151; Smith et al. 2004. Applied and Environmental Microbiology 70:7372–7377.
![Page 9: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/9.jpg)
My Vision: ARG Control
Source of ARGs Technology to TreatARGs
![Page 10: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/10.jpg)
General Research Questions
• What effect does current wastewater treatment operations have on the spread of ARGs?
• How can we reduce ARGs leaving a wastewater treatment facilities?
![Page 11: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/11.jpg)
UntreatedWastewater
BarRack Grit
Chamber
PrimaryClarifier
Cell Recycle
DischargeStream
AerationTank
SecondaryClarifier
SeasonalDisinfection
Where to look?
AnaerobicDigestion(or other)
Farmland
1
23
0
![Page 12: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/12.jpg)
Part 1: Is treated effluent a pertinent source of antibiotic resistance genes?
Photo of treated and raw municipal wastewater from Blue Lake Wastewater Treatment Plant (2003)
![Page 13: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/13.jpg)
Source: LaPara et al. 2015. Multiple discharges of treated municipal wastewater have a small effect on the quantities of numerous antibiotic resistance determinants in the Upper Mississippi River. Environmental Science and Technology 49:11509-11515.
![Page 14: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/14.jpg)
St. Louis River, Duluth-Superior Harbor, WLSSD, and Lake Superior
![Page 15: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/15.jpg)
St. Louis River Lake Superior
Source: LaPara et al. 2011. Tertiary-treated municipal wastewater is a significant point-source of antibiotic resistance genes into Duluth-Superior Harbor. Environmental Science and Technology 45:9543-9549.
Antibiotic resistance genes
![Page 16: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/16.jpg)
Source: LaPara et al. 2015. Multiple discharges of treated municipal wastewater have a small effect on the quantities of numerous antibiotic resistance determinants in the Upper Mississippi River. Environmental Science and Technology 49:11509-11515.
![Page 17: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/17.jpg)
Source: LaPara et al. 2015. Multiple discharges of treated municipal wastewater have a small effect on the quantities of numerous antibiotic resistance determinants in the Upper Mississippi River. Environmental Science and Technology 49:11509-11515.
![Page 18: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/18.jpg)
Part 2: Treatment of Wastewater Solids
• The overwhelming majority of bacteria (antibiotic resistant or otherwise) in sewage end up in sewage sludge
• Treatment technologies vary substantially depending on the end goal – anaerobic digestion at 37°C is most common
![Page 19: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/19.jpg)
Aerobic digestion and air dry slowly reduce ARG levels
Sources: Burch et al. 2013. Environmental Science and Technology 47(17):9965-9971. Burch et al. 2013. Frontiers in Microbiology – Antimicrobials, Resistance and Chemotherapy 4:17. doi: 10.3389/fmicb.2013.00017.
![Page 20: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/20.jpg)
Higher temperature is better at eliminating ARGs during anaerobic digestion
Source: Diehl and LaPara. 2010. Effect of temperature on the fate of genes encoding tetracycline resistance and the integrase of Class 1 integrons within anaerobic and aerobic digesters treating municipal wastewater solids. Environ. Sci. Technol. 44:9128-9133.
= 22°C = 37°C = 46°C = 55°C
Time (d)0 1 2 3 4 5
102
103
104
105tet(X)
Time(d)0 1 2 3 4 5
100
101
102
103
104tet(W) intI1
Time (d)0 1 2 3 4 5
102
103
104
105
106
![Page 21: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/21.jpg)
Part 3: Treated Wastewater Solids are Often Applied to Agricultural Soils
![Page 22: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/22.jpg)
Antibiotic resistance genes decline when wastewater solids are applied to soil
Source: Burch, Sadowsky, and LaPara.2014. Fate of antibiotic resistance genes and class 1 integrons in soil microcosms following the application of treated residual municipal wastewater solids. Environmental Science and Technology 48:5620-5627.
Sandy Soil Silty-Loamy Soil
![Page 23: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/23.jpg)
How do decay rates (t1/2) compare?
Gene Thermophilic Anaerobic Digestion
Aerobic Digestion Air Drying In soils
erm(B) 3.6 6.4 15
intI1 0.28 6.3 31.7 75
sul1 4.6 35.5 55
tet(A) 1.6 4.4 8.8 30
tet(W) 0.96 2.8 7.3 18
tet(X) 0.5 5.7 17.2 40
![Page 24: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/24.jpg)
Take-home messages• Real-time PCR allows us to study the spread
of antibiotic resistance in an entirely new and more mechanistic/quantitative way
• Raw wastewater is bad, bad, bad• Treated wastewater still contains a lot of
antibiotic resistance genes• Thermophilic anaerobic digestion = Great!• Antibiotic resistance genes decay slowly in
soil
![Page 25: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/25.jpg)
What am I really saying?
Wouldn’t it be great to collect a large fraction of antibiotic resistance genes and destroy them?
Our existing infrastructure (sewers & wastewater treatment plants) gets us almost all of the way to that goal!
![Page 26: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/26.jpg)
Significance of My Research• Current efforts to slow the proliferation
of antibiotic resistance are focused on “resistance prevention.”
• I suggest that municipal wastewater treatment can be applied to provide “resistance control.”
• My proposed approach completely supports (does not supplant) on-going resistance prevention efforts
![Page 27: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/27.jpg)
Acknowledgements
• Minnesota Environment and Natural Resources Trust Fund
• National Science Foundation
![Page 28: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/28.jpg)
Acknowledgements
• David Tan• Mi Yan• Tucker Burch• Pat McNamara• Matt Madson• David Diehl• Mike Sadowsky• Kyle Sandberg
• Spencer Borchardt• Kevin Lang• Tim Johnson• Pat McNamara• Matt Madson• David Diehl• Mike Sadowsky• Kyle Sandberg
![Page 29: Dr. Tim Lapara - Stewardship and the Environment - Wastewater Treatment and Antibiotics](https://reader031.vdocument.in/reader031/viewer/2022022411/58f062ed1a28ab67358b4609/html5/thumbnails/29.jpg)
Thank You!