Antibiotic Resistance, Gene Transfer, and Water Quality Patterns Observed in Waterways near CAFO Farms and Wastewater Treatment Facilities
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We examined water quality indicators (pH, temperature, turbidity, total phosphorus, and fecal coliform density) and bacterial antibiotic resistance (prevalence, conjugative transfer, and genetic linkage of resistance elements) at locations impacted by confined animal feeding operations (CAFOs) and compared them to nearby reference sites. Sites located upstream and downstream of two wastewater treatment facilities were also compared. Sites near CAFO farms had poor water quality (elevated total phosphorus and turbidity), while water quality remained relatively good downstream of wastewater treatment plants. High proportions of antibiotic-resistant bacteria were observed at all study sites, and frequent conjugative transfer of resistance was observed in laboratory assays. Out of a total of 830 environmental bacterial isolates, 77.1% were resistant to only ampicillin, while 21.2% were resistant to combinations of antibiotics including ampicillin (A), kanamycin (K), chlorotetracycline (C), oxytetracycline (O), and streptomycin (S). Multi-drug-resistant bacteria were significantly more common at sites impacted by CAFO farms. In conjugation assays, 83.3% of the environmental isolates transferred one or more antibiotic resistance genes to a laboratory strain of Salmonella typhimurium. A subset of multi-drug-resistant (A, C, and O) isolates was screened for specific tetracycline resistance genes and class I and II integrons. None of the screened isolates (n = 22) were positive for integrons, while 13 isolates contained resistance genes for tet (B) and tet (C). Our results indicate that CAFO farms not only impair traditional measures of water quality but may also increase the prevalence of multi-drug-resistant bacteria in natural waters.
KeywordsAntibiotic resistance Conjugation Confined animal feeding operations (CAFOs) Fecal coliforms Water pollution Water quality
We thank Carl F. Marrs and Betsy Foxman, along with their laboratory members, for technical advice and support from the University of Michigan’s Center for Molecular and Clinical Epidemiology of Infectious Diseases (MAC-EPID) program. Janet Kauffman helped locate several appropriate sampling locations. Comments from several anonymous reviewers improved this manuscript.
This work was made possible by a graduate student research award to Bridgette West from the Meta Helwig Scholarship Fund (EMU Biology Department), a sabbatical leave award for Peggy Liggit from Eastern Michigan University, and support from the Eastern Michigan University Center for Aquatic Microbial Ecology.
- American Public Health Association. (1998). Section 9000. Microbial examination. Standard methods for the examination of water and wastewater (20th ed., pp. 56–74). Washington: APHA.Google Scholar
- American Public Health Association. (2005). Section 9260. Detection of pathogenic bacteria. Standard methods for the examination of water and wastewater (21st ed.). Washington: APHA.Google Scholar
- Aminov, R. I., Garrigues-JeanJean, N., & Mackie, R. I. (2001). Molecular ecology of tetracycline resistance: Development and validation of primers for detection of tetracycline resistance genes encoding ribosomal protection proteins. Applied and Environmental Microbiology, 67, 22–32.CrossRefGoogle Scholar
- Aminov, R. I., Chee-Sanford, J. C., Garrigues, N., Teferedegne, B., Krapac, I. J., White, B. A., et al. (2002). Development, validation, and application of PCR primers for detection of tetracycline efflux genes of gram-negative bacteria. Applied and Environmental Microbiology, 68, 1786–1793.CrossRefGoogle Scholar
- Ash, R. J., Mauck, B., & Morgan, M. (2002). Antibiotic resistance of gram-negative bacteria in rivers, United States. Emerging Infectious Diseases, 8, 713–716.Google Scholar
- Bell, J. B., Macrae, W. R., & Elliott, G. E. (1980). Incidence of R factors in coliform, fecal coliform and Salmonella populations of the Red River in Canada. Applied and Environmental Microbiology, 40, 486–491.Google Scholar
- Bell, J. B., Elliott, G. E., & Smith, D. W. (1983). Influence of sewage treatment and urbanization on selection of multiple resistance in fecal coliform populations. Applied and Environmental Microbiology, 46, 227–232.Google Scholar
- Bullas, L. R., & Ryu, J. I. (1983). Salmonella typhimurium LT2 strains which are r−m+ for all three chromosomally located systems of DNA restriction and modification. Journal of Bacteriology, 156, 471–474.Google Scholar
- Burton, G. R. W., & Engelkirk, P. G. (2004). Epidemiology and public health. In J. Goucher, L. Horowitz, & P. C. Williams (Eds.), Microbiology for the health sciences (7th ed., pp. 271–307). Baltimore: Lippincott Williams & Wilkins.Google Scholar
- Chao, K.-K., Chao, C.-C., & Chao, W.-L. (2003). Suitability of the traditional microbial indicators and their enumerating methods in the assessment of fecal pollution of subtropical freshwater environments. Journal of Microbiology, Immunology, and Infection, 36, 288–293.Google Scholar
- Davies, J. E. (1997). Origins, acquisition and dissemination of antibiotic resistance determinants. In D. Chadwick & J. Goode (Eds.), Antibiotic resistance: Origins, evolution, selection and spread (pp. 15–35). New York: Wiley.Google Scholar
- Hall, R. M. (1997). Mobile gene cassettes and integrons: Moving antibiotic resistance genes in gram-negative bacteria. In D. Chadwick & J. Goode (Eds.), Antibiotic resistance: Origins, evolution, selection and spread (pp. 192–205). New York: Wiley.Google Scholar
- Hall, M. L. A., Blok, H. E. M., Donders, A. R. T., Paauw, A., Fluit, A. C., & Verhoef, J. (2003). Multidrug resistance among Enterobacteriaceae is strongly associated with the presence of integrons and is independent of species or isolate origin. The Journal of Infectious Diseases, 187, 251–259.CrossRefGoogle Scholar
- Harwood, V. J., Whitlock, J., & Withington, V. (2000). Classification of antibiotic resistance patterns of indicator bacteria by discriminate analysis: Use in predicting the source of fecal contamination in subtropical waters. Applied and Environmental Microbiology, 66, 3698–3704.CrossRefGoogle Scholar
- Hooper, D. C. (2002). Target modification as a mechanism of antimicrobial resistance. In K. Lewis, A. A. Salyers, H. W. Taber, & R. G. Wax (Eds.), Bacterial resistance to antimicrobials (pp. 161–192). New York: Marcel Dekker.Google Scholar
- Hurd, H. S. (2006). Assessing risks to human health from antibiotic use in food animals. Microbe, 1, 115–119.Google Scholar
- Illinois Pollution Control Board. (2002). Illinois Administrative Code, Title 35, Subtitle C: Water Pollution, Chapter 1, Part 302: Water Quality Standards, Section 302.209: Fecal Coliform (As amended at 12 Ill. Reg. 12082, effective July 11, 1988). Illinois Pollution Control Board, Chicago, Illinois. http://www.ipcb.state.il.us/SLR/IPCBandIEPAEnvironmentalRegulations-Title35.asp. Accessed 21 February 2008.
- Iwane, T., Urase, T., & Yamamoto, K. (2001). Possible impact of treated wastewater discharge on incidence of antibiotic resistant bacteria in river water. Water Science and Technology, 43, 91–99.Google Scholar
- Kauffman, J., & Melmoth, K. (2003). Final report, ECCSCM water monitoring project 2001–2003. Environmentally Concerned Citizens of South Central Michigan, Hudson, MI http://www.nocafos.org/finalreport.htm. Accessed 13 June 2005.
- Khachatourians, G. G. (1998). Agricultural use of antibiotics and the evolution and transfer of antibiotic-resistant bacteria. Canadian Medical Association Journal, 159, 1129–1136.Google Scholar
- Lebaron, P., Roux, V., Lett, M. C., & Baleux, B. (1993). Effects of pili rigidity and energy availability on conjugative plasmid transfer in aquatic environments. Microbial Releases, 2, 127–133.Google Scholar
- Levy, S. B. (1997). Antibiotic resistance: An ecological imbalance. In D. Chadwick & J. Goode (Eds.), Antibiotic resistance: Origins, evolution, selection and spread (pp. 1–14). New York: Wiley.Google Scholar
- Lind, O. T. (1985). Handbook of common methods in limnology (2nd ed.). Dubuque: Hunt.Google Scholar
- Lu, K., Asano, R., & Davies, J. (2004). Antimicrobial resistance gene delivery in animal feeds. Emerging Infectious Diseases, 10, 679–683.Google Scholar
- Mascaretti, O. A. (2003). Bacteria versus antibacterial agents: an integrated approach. Washington: ASM.Google Scholar
- Mazel, D. (2004). Integrons and the origin of antibiotic resistance gene cassettes. ASM News, 70, 520–525.Google Scholar
- McGinnis, S., & Madden, T. L. (2004). BLAST: At the core of a powerful and diverse set of sequence analysis tools. Nucleic Acids Research, 1(32), W20–W25. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&list_uids=15215342&dopt=Citation. Accessed 22 October 2009 (web server issue).CrossRefGoogle Scholar
- Michigan Department of Environmental Quality. (1999). Water quality standards. p A-47 (R323.1062 Microorganisms). Surface Water Quality Division, Michigan Department of Environmental Quality, Lansing, MI. http://www.michigan.gov/deq/0,1607,7-135-3313_3686_3728-12711--,00.html and http://www.deq.state.mi.us/documents/deq-wb-intreport-appendixa.pdf. Accessed 5 January 2007.
- Michigan Department of Environmental Quality. (2003a). Biological surveys of the tributaries in the vicinity of two CAFOs in the St. Joseph and Bean/Tiffin Watersheds Hillsdale and Lenawee counties, Michigan June 10, 11, and 19, 2003. Report #MI/DEQ/WD-03-092. Water Quality Division, Michigan Department of Environmental Quality, Lansing, MI. http://www.michigan.gov/deq/0,1607,7-135-3304-54941--,00.html Accessed 23 October 2006.
- Michigan Department of Environmental Quality. (2003b). Combined sewer overflow and sanitary sewer overflow 2002 & 2003 annual report (January 2002–December 2003). Surface Water Quality Division, Michigan Department of Environmental Quality, Lansing, MI. http://www.deq.state.mi.us/documents/deq-wd-csossoreport03.pdf Accessed 3 June 2005.
- Michigan Department of Environmental Quality. (2004a). Combined sewer overflow and sanitary sewer overflow 2004 annual report. Surface Water Quality Division, Michigan Department of Environmental Quality, Lansing, MI. http://www.deq.state.mi.us/documents/deq-wb-csossoreport04.pdf. Accessed 8 June 2005.
- Michigan Department of Environmental Quality. (2004b). Water quality and pollution control in Michigan: 2004 Sections 303(d) and 305(b) integrated report. Report # MI/DEQWD-04/029. Water Quality Division, Michigan Department of Environmental Quality, Lansing, MI. http://www.michigan.gov/deq/0,1607,7-135-3304-54941--,00.html. Accessed 5 June 2005.
- Michigan Department of Environmental Quality. (2005). Combined sewer overflow and sanitary sewer overflow 2005 annual report. Surface Water Quality Division, Michigan Department of Environmental Quality, Lansing, MI. http://www.deq.state.mi.us/documents/deq-wb-csossoreport05.pdf. Accessed 27 December 2006.
- Mitchell, M. K., & Stapp, W. B. (1997). Field manual for water quality monitoring (11th ed., pp. 25–39). Dubuque: Kendall/Hunt.Google Scholar
- Niemi, M., Sibakov, M., & Niemela, S. (1983). Antibiotic resistance among different species of fecal coliforms isolated from water samples. Applied and Environmental Microbiology, 45, 79–83.Google Scholar
- Ogan, M. T., & Nwiika, D. E. (1993). Studies on the ecology of aquatic bacteria of the lower Niger delta: Multiple antibiotic resistance among the standard plate count organisms. The Journal of Applied Bacteriology, 74, 595–602.Google Scholar
- Quintiliani, R., Jr., Sham, D. F., & Courvalin, P. (1999). Mechanisms of resistance to antimicrobial agents. In P. Murray, E. Baron, M. Pfaller, F. Tenover, & R. Yolken (Eds.), Manual of clinical microbiology (pp. 1505–1525). Washington: ASM.Google Scholar
- Rice, E. W., Messer, J. W., Johnson, C. H., & Reasoner, D. J. (1995). Occurrence of high-level aminoglycoside resistance in environmental isolates of enterococci. Applied and Environmental Microbiology, 61, 374–376.Google Scholar
- Salyers, A. A., Shoemaker, N. B., & Bonheyo, G. T. (2002). The ecology of antibiotic resistance genes. In K. Lewis, A. A. Salyers, H. W. Taber, & R. G. Wax (Eds.), Bacterial resistance to antimicrobials (pp. 1505–1525). New York: Marcel Dekker.Google Scholar
- Sayah, R. S., Kaneene, J. B., Johnson, Y., & Miller, R. (2005). Patterns of antimicrobial resistance observed in Escherichia coli isolates obtained from domestic- and wild-animal fecal samples, human septage, and surface water. Applied and Environmental Microbiology, 71, 1394–1404.CrossRefGoogle Scholar
- Stoner, N. (2005). EPA’s proposed policy on sewage dumping during wet weather conditions. Presented testimony, House Committee on Transportation and Infrastructure's Water Resources and the Environment Subcommittee, April 13, 2005. National Resources Defense Council, New York, NY. http://www.nrdc.org/water/pollution/tns0405.asp. Accessed 21 November 2005.
- Taber, H. W. (2002). Antibiotic permeability. In K. Lewis, A. A. Salyers, H. W. Taber, & R. G. Wax (Eds.), Bacterial resistance to antimicrobials (pp. 193–208). New York: Marcel Dekker.Google Scholar
- Taylor, R. K., Manoil, C., & Mekalanos, J. J. (1989). Broad-host-range vectors for the delivery of TnphoA: Use in genetic analysis of secreted virulence determinants of Vibrio cholerae. Journal of Bacteriology, 171, 1870–1878.Google Scholar
- US Environmental Protection Agency. (2002). United States Code, Title 33, Chapter 26: Federal Water Pollution Control Act. (As amended through P.L. 107-303, November 27, 2002.). US Environmental Protection Agency, Washington, DC. http://www.epa.gov/region5/water/cwa.htm. Accessed 27 December 2006.
- Wetzel, R. G. (2001). Limnology: Lake and river ecosystems (3rd ed.). New York: Academic.Google Scholar
- Wetzel, R. G., & Likens, G. E. (2000). Limnological analyses (3rd ed.). New York: Springer.Google Scholar
- Wiggins, B. A., Andrews, R. W., Conway, R. A., Corr, C. L., Dobratz, E. J., Dougherty, D. P., et al. (1999). Use of antibiotic resistance analysis to identify nonpoint sources of fecal contamination. Applied and Environmental Microbiology, 65, 3483–3486.Google Scholar
- Witte, W. (1997). Impact of antibiotic use in animal feeding on resistance of bacterial pathogens in humans. In D. Chadwick & J. Goode (Eds.), Antibiotic resistance: Origins, evolution, selection and spread (pp. 61–75). New York: Wiley.Google Scholar
- Zar, J. H. (1999). Biostatistical analysis (4th ed.). Upper Saddle River: Prentice Hall.Google Scholar