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 (mg/L)

Water Quality (continued) Ammonia (mg/L)

Water Quality (continued) Total N (mg/L)

Water Quality (continued) Total 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 Image courtesy of

Disk Diffusion: The Concept Image courtesy of

Fleming’s Discovery Image courtesy of

Disk Diffusion: The Procedure Image courtesy of

Disk Diffusion: The Procedure Image courtesy of

Disk Diffusion: The Procedure Image courtesy of

Disk Diffusion: The Procedure Image courtesy of

Disk Diffusion: The Procedure Image courtesy of

Disk Diffusion: The Procedure Image courtesy of

Our specific procedure Collected surface samples Serial dilutions Culture on ColiBlue

ColiBlue media Red = ColiformsBlue = 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 17 mm = Susceptible

Converting Inhibition Zones CLSI standard Example: Ampicillin with 7mm zone of inhibition 17 mm = Susceptible

Results of Testing AmpicillinTetracyclineCiprofloxacinSulfisoxazole Susceptible Intermediate Resistant Total33 34 % Resistant82%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