Microbial Ecology

, Volume 55, Issue 2, pp 184–193 | Cite as

Distribution of Tetracycline and Streptomycin Resistance Genes and Class 1 Integrons in Enterobacteriaceae Isolated from Dairy and Nondairy Farm Soils

  • Velusamy Srinivasan
  • Hyang-Mi Nam
  • Ashish A. Sawant
  • Susan I. Headrick
  • Lien T. Nguyen
  • Stephen P. Oliver
Original Article

Abstract

The prevalence of selected tetracycline and streptomycin resistance genes and class 1 integrons in Enterobacteriaceae (n = 80) isolated from dairy farm soil and nondairy soils was evaluated. Among 56 bacteria isolated from dairy farm soils, 36 (64.3%) were resistant to tetracycline, and 17 (30.4%) were resistant to streptomycin. Lower frequencies of tetracycline (9 of 24 or 37.5%) and streptomycin (1 of 24 or 4.2%) resistance were observed in bacteria isolated from nondairy soils. Bacteria (n = 56) isolated from dairy farm soil had a higher frequency of tetracycline resistance genes including tetM (28.6%), tetA (21.4%), tetW (8.9%), tetB (5.4%), tetS (5.4%), tetG (3.6%), and tetO (1.8%). Among 24 bacteria isolated from nondairy soils, four isolates carried tetM, tetO, tetS, and tetW in different combinations; whereas tetA, tetB, and tetG were not detected. Similarly, a higher prevalence of streptomycin resistance genes including strA (12.5%), strB (12.5%), ant(3″) (12.5), aph(6)-1c (12.5%), aph(3″) (10.8%), and addA (5.4%) was detected in bacteria isolated from dairy farm soils than in nondairy soils. None of the nondairy soil isolates carried aadA gene. Other tetracycline (tetC, tetD, tetE, tetK, tetL, tetQ, and tetT) and streptomycin (aph(6)-1c and ant(6)) resistance genes were not detected in both dairy and nondairy soil isolates. A higher distribution of multiple resistance genes was observed in bacteria isolated from dairy farm soil than in nondairy soil. Among 36 tetracycline- and 17 streptomycin-resistant isolates from dairy farm soils, 11 (30.6%) and 9 (52.9%) isolates carried multiple resistance genes encoding resistance to tetracycline and streptomycin, respectively, which was higher than in bacteria isolated from nondairy soils. One strain each of Citrobacter freundii and C. youngae isolated from dairy farm soils carried class 1 integrons with different inserted gene cassettes. Results of this small study suggest that the presence of multiple resistance genes and class 1 integrons in Enterobacteriaceae in dairy farm soil may act as a reservoir of antimicrobial resistance genes and could play a role in the dissemination of these antimicrobial resistance genes to other commensal and indigenous microbial communities in soil. However, additional longer-term studies conducted in more locations are needed to validate this hypothesis.

References

  1. 1.
    Agerso Y, Sengelov G, Jensen LB (2004) Development of a rapid method for direct detection of tet(M) genes in soil from Danish farmland. Environ Int 30:117–122PubMedCrossRefGoogle Scholar
  2. 2.
    Agerso Y, Sandvang D (2005) Class 1 integrons and tetracycline resistance genes in Alcaligenes, Arthrobacter, and Pseudomonas spp. isolated from pigsties and manured soil. Appl Environ Microbiol 71:794–7947CrossRefGoogle Scholar
  3. 3.
    Aminov RI, Garrigues-Jeanjean N, Mackie RI (2001) Molecular ecology of tetracycline resistance: Development and validation of primers for detection of tetracycline resistance genes encoding ribosomal protection proteins. Appl Environ Microbiol 67:22–32PubMedCrossRefGoogle Scholar
  4. 4.
    Bryan A, Shapir N, Sadowsky MJ (2004) Frequency and distribution of tetracycline resistance genes in generally diverse, nonselected, and nonclinical Escherichia coli strains isolated from diverse human and animal sources. Appl Environ Microbiol 70:2503–2507PubMedCrossRefGoogle Scholar
  5. 5.
    Burgos JM, Ellington BA, Varela MF (2005) Presence of multidrug-resistant enteric bacteria in dairy farm topsoil. J Dairy Sci 88:1391–1398PubMedCrossRefGoogle Scholar
  6. 6.
    Chee-Sanford JC, Aminov RI, Krapac IJ, Garrigues-Jeanjean N, Mackie RI (2001) Occurrence and diversity of tetracycline resistance genes in lagoons and groundwater underlying two swine production facilities. Appl Environ Microbiol 67:1494–1502PubMedCrossRefGoogle Scholar
  7. 7.
    Chopra I, Roberts M (2001) Tetracycline antibiotics: mode of action, application, molecular biology, and epidemiology of bacterial resistance. Microbiol Mol Biol Rev 65:232–260PubMedCrossRefGoogle Scholar
  8. 8.
    Clinical Laboratory Standards Institute (2004) Performance standards for antimicrobial susceptibility testing - fourteenth informational supplement M2-A8 and M7-A6. Wayne, PA, USAGoogle Scholar
  9. 9.
    DeFlaun MF, Levy SB (1989) Genes and their varied hosts, p. I-32. In: Levy, SB, Miller, RV (Ed.) Gene transfer in the environment. McGraw-Hill, New York, NYGoogle Scholar
  10. 10.
    de Freitas JR, Schoenau JJ, Boyetchko SM, Cyrenne SA (2003) Soil microbial populations, community composition, and activity as affected by repeated applications of hog and cattle manure in eastern Saskatchewan. Can J Microbiol 49:538–548PubMedCrossRefGoogle Scholar
  11. 11.
    Fluit AC, Schmitz FJ (1999) Class 1 integrons, gene cassettes, mobility, and epidemiology. Eur J Clin Microbiol Infect Dis 18:761–770PubMedCrossRefGoogle Scholar
  12. 12.
    Food and Drug Administration (2003) National Antimicrobial Resistance Monitoring Systems 2003: Annual Report (FDA, 2003). http://www.fda.gov/cvm/Documents/NARMSRetailMeatRpt2003.pdf
  13. 13.
    Furushita M, Shiba T, Maeda T, Yahata M, Kaneoka A, Takahashi Y, Torri K, Hasegawa T, Ohta M (2003) Similarity of tetracycline resistance genes isolated from fish farm bacteria to those from clinical isolates. Appl Environ Microbiol 69:5339–5342CrossRefGoogle Scholar
  14. 14.
    Gebreyes WA, Altier C (2002) Molecular characterization of multidrug-resistant Salmonella enterica subsp. enterica Serovar Typhimurium isolates from swine. J Clin Microbiol 40:2813–2822PubMedCrossRefGoogle Scholar
  15. 15.
    Goldstein C, Lee MD, Sanchez S, Hudson C, Phillips B, Register B, Grady M, Liebert C, Summers AO, White DG, Maurer JJ (2001) Incidence of class 1 and 2 integrases in clinical and commensal bacteria from livestock, companion animals, and exotics. Antimicrob Agents Chemother 45:723–726PubMedCrossRefGoogle Scholar
  16. 16.
    Guillaume G, Verbrugge D, Chasseur-Libotte ML, Moens W, Collard JM (2000) PCR typing of tetracycline resistance determinants (TetA-E) in Salmonella enterica serotype Hadar and in the microbial community of activated sludges from hospital and urban wastewater treatment facilities in Belgium. FEMS Microbiol Ecol 32:77–85PubMedGoogle Scholar
  17. 17.
    Lanz R, Kuhnert P, Boerlin P (2003) Antimicrobial resistance and resistance gene determinants in clinical Escherichia coli from different animal species in Switzerland. Vet Microbiol 91:73–84PubMedCrossRefGoogle Scholar
  18. 18.
    Lee C, Langlois BE, Dawson KA (1993) Detection of tetracycline resistance determinants in pig isolates from three herds with different histories of antimicrobial exposure. Appl Environ Microbiol 59:1467–1472PubMedGoogle Scholar
  19. 19.
    Levesque C, Piche L, Larose C, Roy PH (1995) PCR mapping of integrons reveals several novel combinations of resistance genes. Antimicrob Agents Chemother 39:185–191PubMedGoogle Scholar
  20. 20.
    Martinez-Freijo P, Fluit AC, Schmitz F-J, Verhoef J, Jones M (1999) Many class I integrons comprise distinct stable structures occurring in different species of Enterobacteriaceae isolated from widespread geographic regions in Europe. Antimicrob Agents Chemother 43:686–689PubMedGoogle Scholar
  21. 21.
    Mazel D, Davies J (1999) Antibiotic resistance in microbes. Cell Mol Life Sci 56:742–754PubMedCrossRefGoogle Scholar
  22. 22.
    McEwen SA, Fedorka-Cray PJ (2002) Antimicrobial use and resistance in animals. Clin Infect Dis 34:S93–S106PubMedCrossRefGoogle Scholar
  23. 23.
    Mukherjee S, Chakraborty R (2006) Incidence of class I integrons in multiple antibiotic-resistant Gram-negative copiotrophic bacteria from the river Torsa in India. Res Microbiol 157:220–226PubMedCrossRefGoogle Scholar
  24. 24.
    Nesvera J, Hochmannova J, Patek M (1998) An integron of class 1 is present on the plasmid pCG4 from Gram-positive bacterium Corynebacterium glutamicum. FEMS Microbiol Lett 169:391–395PubMedCrossRefGoogle Scholar
  25. 25.
    Ng LK, Martin I, Alfa M, Mulvey M (2001) Multiplex PCR for the detection of tetracycline resistant genes. Mol Cell Probes 15:209–215PubMedCrossRefGoogle Scholar
  26. 26.
    Osterblad M, Hakanen A, Manninen R, Leistevuo T, Peltonen R, Meurman O, Huovinen P, Kotilainen P (2000) A between-species comparison of antimicrobial resistance in enterobacteria in fecal flora. Antimicrob Agents Chemother 44:1479–1484PubMedCrossRefGoogle Scholar
  27. 27.
    Osterblad M, Pensala O, Peterzens M, Heleniusc H, Huovinen P (1999) Antimicrobial susceptibility of Enterobacteriaceae isolated from vegetables. J Antimicrob Chemother 43:503–509PubMedCrossRefGoogle Scholar
  28. 28.
    Palmer EL, Teviotdale BL, Jones AL (1997) A relative of the broad-host-range plasmid RSF1010 detected in Erwinia amylovora. Appl Environ Microbiol 63:4604–4607PubMedGoogle Scholar
  29. 29.
    Roberts MC (2003) Tetracycline therapy: update. Clin Infect Dis 36:462–467PubMedCrossRefGoogle Scholar
  30. 30.
    Sengelov G, Agerso Y, Halling-Sorensen B, Baloda SB, Anderson JS, Jensen LB (2003) Bacterial antibiotic resistance levels in Danish farmland as a result of treatment with pig manure slurry. Environ Int 28:587–595PubMedCrossRefGoogle Scholar
  31. 31.
    Shaw KJ, Hare R, Sabatelli FJ, Rizzo M, Cramer CA, Naples L (1991) Correlation between aminoglycoside resistance profiles and DNA hybridization of clinical isolates. Antimicrob Agents Chemother 35:2253–2261PubMedGoogle Scholar
  32. 32.
    Smith MS, Yang RK, Knapp CW, Niu Y, Peak N, Hanfelt MM, Galland JC, Graham DW (2004) Quantification of tetracycline resistance genes in feedlot lagoons by real-time PCR. Appl Environ Microbiol 70:7372–7377PubMedCrossRefGoogle Scholar
  33. 33.
    Smith DL, Harris AD, Johnson JA, Silbregeld EK, Morris JG Jr (2002) Animal antibiotic use has an early but important impact on the emergence of antibiotic resistance in human commensal bacteria. Proc Natl Acad Sci USA 99:6434–6439PubMedCrossRefGoogle Scholar
  34. 34.
    Sundin GW, Bender CL (1996) Molecular analysis of closely-related copper- and streptomycin-resistance plasmids in Pseudomonas syringae pv. syringae. Plasmid 35:98–107PubMedCrossRefGoogle Scholar
  35. 35.
    Sundin GW, Monks DE, Bender CL (1995) Distribution of the streptomycin-resistance transposons Tn5393 among phylloplane and soil bacteria from managed agricultural habitats. Can J Microbiol 41:792–799PubMedCrossRefGoogle Scholar
  36. 36.
    van Overbeek LS, Wellington EMH, Egan S, Smalla K, Heuer H, Collard JM, Guillaume G, Karagouni AD, Nikolakopoulou TL, van Elsas JD (2002) Prevalence of streptomycin-resistance genes in bacterial populations in European habitats. FEMS Microbiol Ecol 42:277–288PubMedGoogle Scholar
  37. 37.
    Villedieu A, Diaz-Torres ML, Hunt N, McNab R, Spratt DA, Wilson M, Mullany P (2003) Prevalence of tetracycline resistance genes in oral bacteria. Antimicrob Agents Chemother 47:878–882PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Velusamy Srinivasan
    • 1
  • Hyang-Mi Nam
    • 1
  • Ashish A. Sawant
    • 1
  • Susan I. Headrick
    • 1
  • Lien T. Nguyen
    • 1
  • Stephen P. Oliver
    • 1
  1. 1.Department of Animal Science and the Food Safety Center of ExcellenceThe University of TennesseeKnoxvilleUSA

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