Quantification of tetracycline resistance genes in feedlot lagoons by real-time PCR

M.S. Smith, R.K. Yang, C.W. Knapp, Y.F. Niu, N. Peak, M.M. Hanfelt, J.C. Galland, D.W. Graham

Research output: Contribution to journalArticle

134 Citations (Scopus)

Abstract

A new real-time PCR method is presented that detects and quantifies three tetracycline resistance (Tc-r) genes [tet(O), tet(W), and tet(Q)] in mixed microbial communities resident in feedlot lagoon wastewater. Tc-r gene real-time TaqMan primer-probe sets were developed and optimized to quantify the Tc-r genes present in seven different cattle feedlot lagoons, to validate the method, and to assess whether resistance gene concentrations correlate with free-tetracycline levels in lagoon waters. The method proved to be sensitive across a wide range of gene concentrations and provided consistent and reproducible results from complex lagoon water samples. The log,0 of the sum of the three resistance gene concentrations was correlated with free-tetracycline levels (r(2) = 0.50, P<0.001; n = 18), with the geometric means of individual resistance concentrations ranging from 4- to 8.3-fold greater in lagoon samples with above-median tetracycline levels (>1.95 mug/liter by enzyme-linked immunosorbent assay techniques) than in below-median lagoon samples. Of the three Tc-r genes tested, tet(W) and tet(Q) were more commonly found in lagoon water samples. Successful development of this real-time PCR assay will permit other studies quantifying Tc-r gene numbers in environmental and other samples.
LanguageEnglish
Pages7372-7377
Number of pages5
JournalApplied and Environmental Microbiology
Volume70
Issue number12
DOIs
Publication statusPublished - Dec 2004

Fingerprint

Tetracycline Resistance
feedlots
tetracycline
Real-Time Polymerase Chain Reaction
lagoon
quantitative polymerase chain reaction
gene
Genes
genes
Tetracycline
Water
sampling
Immunosorbent Techniques
assay
Waste Water
methodology
water
microbial communities
wastewater
cattle

Keywords

  • quantification
  • tetracycline resistance genes
  • feedlot lagoons
  • real-time PCR

Cite this

Smith, M.S. ; Yang, R.K. ; Knapp, C.W. ; Niu, Y.F. ; Peak, N. ; Hanfelt, M.M. ; Galland, J.C. ; Graham, D.W. / Quantification of tetracycline resistance genes in feedlot lagoons by real-time PCR. In: Applied and Environmental Microbiology. 2004 ; Vol. 70, No. 12. pp. 7372-7377.
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Smith, MS, Yang, RK, Knapp, CW, Niu, YF, Peak, N, Hanfelt, MM, Galland, JC & Graham, DW 2004, 'Quantification of tetracycline resistance genes in feedlot lagoons by real-time PCR' Applied and Environmental Microbiology, vol. 70, no. 12, pp. 7372-7377. https://doi.org/10.1128/AEM.70.12.7372-7377.2004

Quantification of tetracycline resistance genes in feedlot lagoons by real-time PCR. / Smith, M.S.; Yang, R.K.; Knapp, C.W.; Niu, Y.F.; Peak, N.; Hanfelt, M.M.; Galland, J.C.; Graham, D.W.

In: Applied and Environmental Microbiology, Vol. 70, No. 12, 12.2004, p. 7372-7377.

Research output: Contribution to journalArticle

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T1 - Quantification of tetracycline resistance genes in feedlot lagoons by real-time PCR

AU - Smith, M.S.

AU - Yang, R.K.

AU - Knapp, C.W.

AU - Niu, Y.F.

AU - Peak, N.

AU - Hanfelt, M.M.

AU - Galland, J.C.

AU - Graham, D.W.

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KW - feedlot lagoons

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JF - Applied and Environmental Microbiology

SN - 0099-2240

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ER -