Hierarchal clustering yields insight into multidrug-resistant bacteria isolated from a cattle feedlot wastewater treatment system

  • Michael A. Jahne
  • Shane W. Rogers
  • Ivan P. Ramler
  • Edith Holder
  • Gina Hayes


Forty-two percent of Escherichia coli and 58 % of Enterococcus spp. isolated from cattle feedlot runoff and associated infiltration basin and constructed wetland treatment system were resistant to at least one antibiotic of clinical importance; a high level of multidrug resistance (22 % of E. coli and 37 % of Enterococcus spp.) was observed. Hierarchical clustering revealed a closely associated resistance cluster among drug-resistant E. coli isolates that included cephalosporins (ceftiofur, cefoxitin, and ceftriaxone), aminoglycosides (gentamycin, kanamycin, and amikacin), and quinolone nalidixic acid; antibiotics from these classes were used at the study site, and cross-resistance may be associated with transferrable multiple-resistance elements. For Enterococcus spp., co-resistance among vancomycin, linezolid, and daptomycin was common; these antibiotics are reserved for complicated clinical infections and have not been approved for animal use. Vancomycin resistance (n = 49) only occurred when isolates were resistant to linezolid, daptomycin, and all four of the MLSB (macrolide-lincosamide-streptogramin B) antibiotics tested (tylosin, erythromycin, lincomycin, and quinipristin/dalfopristin). This suggests that developing co-resistance to MLSB antibiotics along with cyclic lipopeptides and oxazolidinones may result in resistance to vancomycin as well. Effects of the treatment system on antibiotic resistance were pronounced during periods of no rainfall and low flow (long residence time). Increased hydraulic loading (short residence time) under the influence of rain caused antibiotic-resistant bacteria to be flushed through the treatment system. This presents concern for environmental discharge of multidrug-resistant organisms relevant to public health.


Antibiotic resistance Hierarchal clustering Animal feeding operations Fecal indicator bacteria Wastewater Vancomycin 



ATP-binding cassette


extended-spectrum beta-lactamases


fecal indicator bacteria


U.S. Food and Drug Administration


Jaccard similarity


macrolide-lincosamide-streptogramin B


most probable number


methicillin-resistant Staphylococcus aureus


undecaprenyl phosphate


undecaprenyl pyrophosphate


U.S. Environmental Protection Agency


vancomycin-resistant Enterococcus faecium



This project was supported by the National Research Initiative of the USDA Cooperative State Research, Education, and Extension Service, grant number 2007-35102-18614. The U.S. Environmental Protection Agency, through its Office of Research and Development, also funded and collaborated in the research described herein. Any opinions expressed in this paper are those of the authors and do not necessarily reflect the official positions and policies of the USEPA. Any mention of trade names or commercial products does not constitute endorsement or recommendation for use. This material is based, in part, upon work supported by the National Science Foundation under Grant No. 0959713. The authors thank Dr. Robert Burns of Iowa State University for installation, calibration, and maintenance of the sampling equipment and collection and shipment of manure and water samples. The authors also thank the landowner who graciously allowed access and collection of samples at the study site.


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Copyright information

© Springer International Publishing Switzerland (outside the USA) 2014

Authors and Affiliations

  • Michael A. Jahne
    • 1
  • Shane W. Rogers
    • 2
    • 6
  • Ivan P. Ramler
    • 3
  • Edith Holder
    • 4
  • Gina Hayes
    • 5
    • 7
  1. 1.Institute for a Sustainable EnvironmentClarkson UniversityPotsdamUSA
  2. 2.National Risk Management Research LaboratoryU.S. Environmental Protection AgencyCincinnatiUSA
  3. 3.Department of Mathematics, Computer Science, and StatisticsSt. Lawrence UniversityCantonUSA
  4. 4.Pegasus Technical ServicesCincinnatiUSA
  5. 5.Institute for the Environment and SustainabilityMiami UniversityOxfordUSA
  6. 6.Department of Civil and Environmental EngineeringClarkson UniversityPotsdamUSA
  7. 7.Ohio EPALazarus Government CenterColumbusUSA

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