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Antibiotic Resistant Escherichia Coli in Non-Tidal Wetlands of Maryland

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Title: Antibiotic Resistant Escherichia Coli in Non-Tidal Wetlands of Maryland


1
Antibiotic Resistant Escherichia Coli in
Non-Tidal Wetlands of Maryland
  • Speakers Neil Agarwal Robert Vocke III
  • University of Maryland, College Park

2
Establishing a Baseline
  • MDE
  • Clean Water Act
  • Antibiotics

3
Site Overview
  • Non-tidal, mitigation sites
  • 3 plots per site

4
Data Collection
  • Vegetation
  • Soils
  • Land Use
  • Water Quality
  • Antibiotic Resistance

5
Our focus today
  • Water quality data
  • Why Antibiotic Resistance?
  • The Concept of Measuring Resistance
  • Preliminary Results

6
Water Quality Results
Nitrate (mg/L)
7
Water Quality (continued)
Ammonia (mg/L)
8
Water Quality (continued)
Total N (mg/L)
9
Water Quality (continued)
Total P (mg/L)
10
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

11
Disk Diffusion The Concept
Image courtesy of http//biology.clc.uc.edu/fankha
user/Labs/Microbiology
Image courtesy of http//jyi.org/articleimages/555
/
12
Disk Diffusion The Concept
Image courtesy of http//www.wings-china.com/showi
nfo.asp?typeid24xxid554LanguageEn
13
Flemings Discovery
Image courtesy of http//www.accessexcellence.org/
AE/AEC/CC/s5.html
14
Disk Diffusion The Procedure
Image courtesy of http//jyi.org/articleimages/555
/
15
Disk Diffusion The Procedure
Image courtesy of http//jyi.org/articleimages/555
/
16
Disk Diffusion The Procedure
Image courtesy of http//jyi.org/articleimages/555
/
17
Disk Diffusion The Procedure
Image courtesy of http//jyi.org/articleimages/555
/
18
Disk Diffusion The Procedure
Image courtesy of http//jyi.org/articleimages/555
/
19
Disk Diffusion The Procedure
Image courtesy of http//jyi.org/articleimages/555
/
20
Our specific procedure
  • Collected surface samples
  • Serial dilutions
  • Culture on ColiBlue

21
ColiBlue media
Red Coliforms
Blue E. Coli
22
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

23
Converting Inhibition Zones
  • CLSI standard
  • Example
  • Ampicillin with 7mm zone of inhibition

lt13 mm Resistant 14-16 mm Intermediate
gt17 mm Susceptible
24
Converting Inhibition Zones
  • CLSI standard
  • Example
  • Ampicillin with 7mm zone of inhibition

lt13 mm Resistant 14-16 mm Intermediate
gt17 mm Susceptible
25
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
26
Whats next?
  • Find correlations to health
  • Test upstream downstream
  • Effect on nutrient cycling

27
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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