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Antibiotic Resistant Escherichia Coli in Non-Tidal Wetlands of Maryland

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.

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Antibiotic Resistant Escherichia Coli in Non-Tidal Wetlands of Maryland

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  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/fankhauser/Labs/Microbiology Image courtesy of http://jyi.org/articleimages/555/

  12. Disk Diffusion: The Concept Image courtesy of http://www.wings-china.com/showinfo.asp?typeid=24&xxid=554&Language=En

  13. Fleming’s 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 <13 mm = Resistant 14-16 mm = Intermediate >17 mm = Susceptible

  24. Converting Inhibition Zones • CLSI standard • Example: • Ampicillin with 7mm zone of inhibition <13 mm = Resistant 14-16 mm = Intermediate >17 mm = Susceptible

  25. Results of Testing

  26. What’s 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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