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Applied and Environmental Microbiology, January 2009, p. 203-211, Vol. 75, No. 1
0099-2240/09/$08.00+0     doi:10.1128/AEM.01254-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Quantifying Nonspecific TEM β-Lactamase (blaTEM) Genes in a Wastewater Stream{triangledown}

Karen L. Lachmayr,1* Lee J. Kerkhof,2 A. Gregory DiRienzo,3 Colleen M. Cavanaugh,1 and Timothy E. Ford4

The Biological Laboratories, Harvard University, Cambridge, Massachusetts 02138,1 Institute of Marine and Coastal Sciences, Rutgers University, New Brunswick, New Jersey 08901,2 Department of Epidemiology and Biostatistics, School of Public Health, University at Albany—SUNY, Rensselaer, New York 12144,3 University of New England, Biddeford, Maine 040054

Received 5 June 2008/ Accepted 24 October 2008

To control the antibiotic resistance epidemic, it is necessary to understand the distribution of genetic material encoding antibiotic resistance in the environment and how anthropogenic inputs, such as wastewater, affect this distribution. Approximately two-thirds of antibiotics administered to humans are β-lactams, for which the predominant bacterial resistance mechanism is hydrolysis by β-lactamases. Of the β-lactamases, the TEM family is of overriding significance with regard to diversity, prevalence, and distribution. This paper describes the design of DNA probes universal for all known TEM β-lactamase genes and the application of a quantitative PCR assay (also known as Taqman) to quantify these genes in environmental samples. The primer set was used to study whether sewage, both treated and untreated, contributes to the spread of these genes in receiving waters. It was found that while modern sewage treatment technologies reduce the concentrations of these antibiotic resistance genes, the ratio of blaTEM genes to 16S rRNA genes increases with treatment, suggesting that bacteria harboring blaTEM are more likely to survive the treatment process. Thus, β-lactamase genes are being introduced into the environment in significantly higher concentrations than occur naturally, creating reservoirs of increased resistance potential.

* Corresponding author. Mailing address: The Biological Laboratories, Harvard University, 16 Divinity Ave., Cambridge, MA 02138. Phone: (617) 496-6940. Fax: (617) 496-6933. E-mail: lachmayr{at}post.harvard.edu

{triangledown} Published ahead of print on 7 November 2008.

Applied and Environmental Microbiology, January 2009, p. 203-211, Vol. 75, No. 1
0099-2240/09/$08.00+0     doi:10.1128/AEM.01254-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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