This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Yu, Z.
Right arrow Articles by Morrison, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yu, Z.
Right arrow Articles by Morrison, M.
Agricola
Right arrow Articles by Yu, Z.
Right arrow Articles by Morrison, M.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, November 2005, p. 6926-6933, Vol. 71, No. 11
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.11.6926-6933.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Development and Application of Real-Time PCR Assays for Quantification of Genes Encoding Tetracycline Resistance

Zhongtang Yu,1* Frederick C. Michel Jr,2 Glenn Hansen,3 Thomas Wittum,3 and Mark Morrison1

The MAPLE Research Initiative, Department of Animal Sciences,1 Department of Food, Agricultural and Biological Engineering,2 Department of Veterinary Preventive Medicine, The Ohio State University, Columbus, Ohio 432103

Received 25 March 2005/ Accepted 30 June 2005

We report here the development, validation, and use of three real-time PCR assays to quantify the abundance of the following three groups of tetracycline resistance genes: tet(A) and tet(C); tet(G); and tet genes encoding ribosomal protection proteins, including tet(M), tet(O), tetB(P), tet(Q), tet(S), tet(T), and tet(W). The assays were validated using known numbers of sample-derived tet gene templates added to microbiome DNA. These assays are both precise and accurate over at least 6 log tet gene copies. New tet gene variants were also identified from cloned tet amplicons as part of this study. The utility of these real-time PCR assays was demonstrated by quantifying the three tet gene groups present in bovine and swine manures, composts of swine manure, lagoons of hog house effluent, and samples from an Ekokan upflow biofilter system treating hog house effluent. The bovine manures were found to contain fewer copies of all three groups of tet genes than the swine manures. The composts of swine manures had substantially reduced tet gene abundance (up to 6 log), while lagoon storage or the upflow biofilter had little effect on tet gene abundance. These results suggest that the method of manure storage and treatment may have a substantial impact on the persistence and dissemination of tet genes in agricultural environments. These real-time PCR assays provide rapid, quantitative, cultivation-independent measurements of 10 major classes of tet genes, which should be useful for ecological studies of antibiotic resistance.

* Corresponding author. Mailing address: Department of Animal Sciences, The Ohio State University, 2027 Coffey Road, Columbus, OH 43210. Phone: (614) 292-3057. Fax: (614) 292-7116. E-mail: yu.226{at}osu.edu.

Applied and Environmental Microbiology, November 2005, p. 6926-6933, Vol. 71, No. 11
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.11.6926-6933.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Chee-Sanford, J. C., Mackie, R. I., Koike, S., Krapac, I. G., Lin, Y.-F., Yannarell, A. C., Maxwell, S., Aminov, R. I. (2009). Fate and Transport of Antibiotic Residues and Antibiotic Resistance Genes following Land Application of Manure Waste. J. Environ. Qual. 38: 1086-1108 [Abstract] [Full Text]  
  • Sharma, R., Larney, F. J., Chen, J., Yanke, L. J., Morrison, M., Topp, E., McAllister, T. A., Yu, Z. (2009). Selected Antimicrobial Resistance during Composting of Manure from Cattle Administered Sub-Therapeutic Antimicrobials. J. Environ. Qual. 38: 567-575 [Abstract] [Full Text]  
  • Lachmayr, K. L., Kerkhof, L. J., DiRienzo, A. G., Cavanaugh, C. M., Ford, T. E. (2009). Quantifying Nonspecific TEM {beta}-Lactamase (blaTEM) Genes in a Wastewater Stream. Appl. Environ. Microbiol. 75: 203-211 [Abstract] [Full Text]  
  • Chen, J., Fluharty, F. L., St-Pierre, N., Morrison, M., Yu, Z. (2008). Technical note: Occurrence in fecal microbiota of genes conferring resistance to both macrolide-lincosamide-streptogramin B and tetracyclines concomitant with feeding of beef cattle with tylosin. J ANIM SCI 86: 2385-2391 [Abstract] [Full Text]  
  • Yu, Z., Garcia-Gonzalez, R., Schanbacher, F. L., Morrison, M. (2008). Evaluations of Different Hypervariable Regions of Archaeal 16S rRNA Genes in Profiling of Methanogens by Archaea-Specific PCR and Denaturing Gradient Gel Electrophoresis. Appl. Environ. Microbiol. 74: 889-893 [Abstract] [Full Text]  
  • Storteboom, H. N., Kim, S.-C., Doesken, K. C., Carlson, K. H., Davis, J. G., Pruden, A. (2007). Response of Antibiotics and Resistance Genes to High-Intensity and Low-Intensity Manure Management. J. Environ. Qual. 36: 1695-1703 [Abstract] [Full Text]  
  • He, S., Gall, D. L., McMahon, K. D. (2007). "Candidatus Accumulibacter" Population Structure in Enhanced Biological Phosphorus Removal Sludges as Revealed by Polyphosphate Kinase Genes. Appl. Environ. Microbiol. 73: 5865-5874 [Abstract] [Full Text]  
  • Bibbal, D., Dupouy, V., Ferre, J. P., Toutain, P. L., Fayet, O., Prere, M. F., Bousquet-Melou, A. (2007). Impact of Three Ampicillin Dosage Regimens on Selection of Ampicillin Resistance in Enterobacteriaceae and Excretion of blaTEM Genes in Swine Feces. Appl. Environ. Microbiol. 73: 4785-4790 [Abstract] [Full Text]  
  • Chen, J., Yu, Z., Michel, F. C. Jr., Wittum, T., Morrison, M. (2007). Development and Application of Real-Time PCR Assays for Quantification of erm Genes Conferring Resistance to Macrolides-Lincosamides-Streptogramin B in Livestock Manure and Manure Management Systems. Appl. Environ. Microbiol. 73: 4407-4416 [Abstract] [Full Text]  
  • Kaneene, J. B., Miller, R., Sayah, R., Johnson, Y. J., Gilliland, D., Gardiner, J. C. (2007). Considerations When Using Discriminant Function Analysis of Antimicrobial Resistance Profiles To Identify Sources of Fecal Contamination of Surface Water in Michigan. Appl. Environ. Microbiol. 73: 2878-2890 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»