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Antibiotic Resistant Escherichia Coli in Non-Tidal Wetlands of Maryland Speakers: Neil Agarwal & Robert Vocke III University of Maryland, College Park
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Antibiotic Resistant Escherichia Coli in Non-Tidal Wetlands of Maryland
Speakers: Neil Agarwal & Robert Vocke III
University of Maryland, College Park
Establishing a Baseline
MDE
Clean Water Act
Antibiotics
Site Overview
Non-tidal, mitigation sites 3 plots per site
Data Collection
Vegetation
Soils
Land Use
Water Quality
Antibiotic Resistance
Our focus today…
Water quality data
Why Antibiotic Resistance?
The Concept of Measuring Resistance
Preliminary Results
Water Quality Results
Nitrate
0.050
1.246
0.000
0.200
0.400
0.600
0.800
1.000
1.200
1.400
1.600
1.800
Surface Subsurface
Nit
rate
(m
g/L
)
Water Quality (continued)
Ammonia
0.924
0.221
0.000
0.200
0.400
0.600
0.800
1.000
1.200
Surface Subsurface
Am
mo
nia
(m
g/L
)
Water Quality (continued)
Total Nitrogen
4.4904.267
0.000
1.000
2.000
3.000
4.000
5.000
6.000
Surface Subsurface
To
tal
N (
mg
/L)
Water Quality (continued)
Total Phosphorus
2.300
0.487
0.000
0.500
1.000
1.500
2.000
2.500
3.000
3.500
4.000
Surface Subsurface
To
tal
P (
mg
/L)
Why look at antibiotic resistance? The pharmaceutical problem
Antibiotics and resistance Testing for antibiotic resistance is cost
effective Questions to be asked:
Prevalence and temporal changes Correlation to other aspects of health Resistance ‘sinks’ Nutrient Cycling
Disk Diffusion: The Concept
Image courtesy of http://biology.clc.uc.edu/fankhauser/Labs/Microbiology
Image courtesy of http://jyi.org/articleimages/555/
Disk Diffusion: The Concept
Image courtesy of http://www.wings-china.com/showinfo.asp?typeid=24&xxid=554&Language=En
Fleming’s Discovery
Image courtesy of http://www.accessexcellence.org/AE/AEC/CC/s5.html
Disk Diffusion: The Procedure
Image courtesy of http://jyi.org/articleimages/555/
Disk Diffusion: The Procedure
Image courtesy of http://jyi.org/articleimages/555/
Disk Diffusion: The Procedure
Image courtesy of http://jyi.org/articleimages/555/
Disk Diffusion: The Procedure
Image courtesy of http://jyi.org/articleimages/555/
Disk Diffusion: The Procedure
Image courtesy of http://jyi.org/articleimages/555/
Disk Diffusion: The Procedure
Image courtesy of http://jyi.org/articleimages/555/
Our specific procedure
Collected surface samples Serial dilutions Culture on ColiBlue
ColiBlue media
Red = Coliforms Blue = E. Coli
Our specific procedure
Collected surface samples Serial dilutions Culture on ColiBlue Isolate a single E. coli colony Disk diffusion using 5 antibiotics:
Ampicillin Erythromycin Tetracycline Sulfisoxazole Ciprofloxacin Negative Control: Vancomycin
Converting Inhibition Zones
CLSI standard Example: Ampicillin with 7mm
zone of inhibition
<13 mm = Resistant 14-16 mm = Intermediate >17 mm = Susceptible
Converting Inhibition Zones
CLSI standard Example: Ampicillin with 7mm
zone of inhibition
<13 mm = Resistant 14-16 mm = Intermediate >17 mm = Susceptible
Results of Testing
Ampicillin Tetracycline Ciprofloxacin Sulfisoxazole
Susceptible 1 2 32 11
Intermediate 5 13 2 12
Resistant 27 18 0 11
Total 33 33 34 34
% Resistant 82% 55% 0% 32%
What’s next?
Find correlations to health Test upstream & downstream Effect on nutrient
cycling
Antibiotic Resistant Escherichia Coli in Non-Tidal Wetlands of Maryland
Speakers: Neil Agarwal & Robert Vocke III
University of Maryland, College Park
