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Published online ahead of print on 23 April 2009 as doi:10.1099/mic.0.028233-0
Microbiology 2009;155:2306.

Microbiology (2009), DOI 10.1099/mic.0.028233-0
© 2009 Society for General Microbiology
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Microbiology 0 (2009), mic.0.028233; DOI  10.1099/mic.0.028233-0
© 2009 Society for General Microbiology

Detection of 140 clinically relevant antibiotic resistance genes in the plasmid metagenome of wastewater treatment plant bacteria showing reduced susceptibility to selected antibiotics

Rafael Szczepanowski1, Burkhard Linke1, Irene Krahn1, Karl-Heinz Gartemann1, Tim Götzkow1, Wolfgang Eichler2, Alfred Pühler1 and Andreas Schlüter1,3

1 Bielefeld University;
2 Landesamt für Natur, Umwelt und Verbraucherschutz NRW

ABSTRACT

To detect plasmid-borne antibiotic resistance genes in wastewater treatment plant (WWTP) bacteria, 192 resistance gene-specific PCR-primer pairs were designed and synthesised. Subsequent PCR analyses on total plasmid-DNA preparations obtained from bacteria of activated sludge or the WWTP's final effluents led to the identification of, respectively, 140 and 123 different resistance gene-specific amplicons. Among the detected genes are aminoglycoside, beta-lactam, chloramphenicol, fluoroquinolone, macrolide, rifampicin, tetracycline, trimethoprim and sulphonamide resistance genes as well as multidrug efflux and small multidrug resistance genes. Some of these genes were only recently described from clinical isolates demonstrating genetic exchange between clinical and WWTP bacteria. Sequencing of selected resistance gene-specific amplicons confirmed their identity or revealed that the amplicon nucleotide sequence is very similar to a gene closely related to the reference gene used for primer design. These results demonstrate that wastewater treatment plant bacteria are a reservoir for various resistance genes. Moreover, detection of about 64% of the 192 reference resistance genes in bacteria obtained from the WWTP's final effluents, indicates that these resistance determinants might be further disseminated in habitats downstream the sewage plant.

3 E-mail: andreas.schlueter{at}genetik.uni-bielefeld.de






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