Monitoring and source tracking of tetracycline resistance genes in lagoons and groundwater adjacent to swine production facilities over a 3-year period

S. Koike, I. G. Krapac, H. D. Oliver, A. C. Yannarell, J. C. Chee-Sanford, Rustam Aminov, R. I. Mackie

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Abstract

To monitor the dissemination of resistance genes into the environment, we determined the occurrence of tetracycline resistance (Tc-r) genes in groundwater underlying two swine confinement operations. Monitoring well networks (16 wells at site A and 6 wells at site C) were established around the lagoons at each facility. Groundwater (n = 124) and lagoon (n = 12) samples were collected from the two sites at six sampling times from 2000 through 2003. Total DNA was extracted, and PCR was used to detect seven TCr genes [tet(M), tet(O), tet(Q), tet(W), tet(C), tet(H), and tet(Z)]. The concentration of Tc-r genes was quantified by real-time quantitative PCR. To confirm the Tc-r gene source in groundwater, comparative analysis of tet(W) gene sequences was performed on groundwater and lagoon samples. All seven Tc-r genes were continually detected in groundwater during the 3-year monitoring period at both sites. At site A, elevated detection frequency and concentration of Tc-r genes were observed in the wells located down-gradient of the lagoon. Comparative analysis of tet(W) sequences revealed that the impacted groundwater contained gene sequences almost identical (99.8% identity) to those in the lagoon, but these genes were not found in background libraries. Novel sequence clusters and unique indigenous resistance gene pools were also found in the groundwater. Thus, antibiotic resistance genes in groundwater are affected by swine manure, but they are also part of the indigenous gene pool.
Original languageEnglish
JournalAPPLIED AND ENVIRONMENTAL MICROBIOLOGY
Volume73
Issue number15
Pages (from-to)4813-4823
Number of pages11
ISSN0099-2240
DOIs
Publication statusPublished - 2007
Externally publishedYes

Keywords

  • Animal Husbandry
  • Animals
  • DNA, Bacterial
  • Fresh Water
  • Genes, Bacterial
  • Manure
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • RNA, Ribosomal, 16S
  • Sequence Analysis, DNA
  • Swine
  • Tetracycline Resistance
  • Antibiotics
  • DNA
  • Groundwater
  • Sewage lagoons
  • Wells
  • Genes
  • ground water
  • tetracycline
  • antibiotic resistance
  • comparative study
  • groundwater
  • lagoon
  • monitoring
  • polymerase chain reaction
  • technetium
  • ungulate
  • article
  • DNA extraction
  • environmental monitoring
  • gene
  • gene cluster
  • gene pool
  • gene sequence
  • manure
  • nonhuman
  • nucleotide sequence
  • quantitative analysis
  • real time polymerase chain reaction
  • swine
  • teteO gene
  • tetH gene
  • tetM gene
  • tetQ gene
  • tetW gene
  • tetZ gene
  • water sampling
  • Suidae
  • Comparative analysis
  • Gene pool
  • Well networks
  • TETRACYCLINE
  • T
  • X
  • BIOTECHNOLOGY
  • MICROBIOLOGY
  • REAL-TIME PCR
  • MEGASPHAERA-ELSDENII STRAINS
  • ANTIBIOTIC-RESISTANCE
  • FECAL POLLUTION
  • RIBOSOMAL-RNA
  • BACTERIA
  • QUANTIFICATION
  • STREPTOCOCCI
  • VALIDATION
  • INFERENCE
  • groundwater adjacent
  • swine manure
  • Artiodactyla Mammalia Vertebrata Chordata Animalia (Animals, Artiodactyls, Chordates, Mammals, Nonhuman Vertebrates, Nonhuman Mammals, Vertebrates) - Suidae [85740] swine common
  • swine tet(C) gene [Suidae]
  • swine tet(H) gene [Suidae]
  • swine tet(M) gene [Suidae]
  • swine tet(O) gene [Suidae]
  • swine tet(Q) gene [Suidae]
  • swine tet(W) gene [Suidae]
  • swine tet(Z) gene [Suidae]
  • swine tetracycline resistance gene [Suidae]
  • total DNA
  • 03502, Genetics - General
  • 03506, Genetics - Animal
  • 07502, Ecology: environmental biology - General and methods
  • 07508, Ecology: environmental biology - Animal
  • 26502, Animal production - General and methods
  • Agriculture
  • Biochemistry and Molecular Biophysics
  • Environmental Sciences
  • real-time quantitative polymerase chain reaction laboratory techniques, genetic techniques
  • Ecology
  • Molecular Genetics

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