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Abstract Antibiotic Resistance Transfer in Agriculture Investigation into Antibiotic Resistance in Bacteria in Agricultural Settings Hypothesis 1 - Resistance due to lateral gene transfer Resistance highly specific to antibiotics used Resistance genes may be carried on plasmids ?-Lactamase gene may resemble class A TEM bla found in enterics Hypothesis 2 (null) - Resistance evolved in soil microorganisms Broader resistance to variety of antibiotics encountered in soil over time Resistance may be plasmid or chromosomally encoded Class B Metallo- ?-Lactamase observed in Caulobacter may be present Water Collection Sites Meat Cattle farm in Swinton, MO. Antibiotic use reported as penicillin only. 100 ml water samples were taken from 3 ponds and a creek adjacent to the farm ( ) UTM 16 coordinates shown for map datum NAD 27 in CONUS. ArcMap used to plot the points onto the topographic map. Isolation Approach Ampicillin resistance Plated on APT agar w/ 50 ?g/ml ampicillin Single colony taken from each plate with growth, unless additional morphotypes present Screened for cefinase activity Non-coliform status (accepted if one of the following are true) Gram positive No lactose fermentation on EMB No gas from lactose broth Organisms Documentation of Resistance MIC of Ampicillin for isolates was determined with Etest? strips (upper left) Result: 14 of the isolates had a MIC of greater than 256 μg/ml. CPB30 (96μg/ml) and CPC32 (128 μg/ml) were slightly lower. Kirby-Bauer Agar diffusion tests (upper right) were used to test for resistance to ?-lactam (Oxacillin, Cefaclor, Cefazolin, Cefotaxime, Imipenem, Carbenicillin )and non- ?-lactam antibiotics (Levaquin, tetracycline, Polymyxin B, Erythromycin, Kanamycin, Streptomycin, Rifampin, Novobiocin). Result 1: All of the isolates were resistant to at least 1 non-?-lactam antibiotic, and 14 were resistant to 2 or more. Result 2: None of the isolates showed resistance to imipenem, suggesting no metallo- ?-lactamase activity