Distribution of specific tetracycline and erythromycin resistance genes in environmental samples assessed by macroarray detection

A macroarray system was developed to screen environmental samples for the presence of specific tetracycline (TcR) and erythromycin (ermR) resistance genes. The macroarray was loaded with polymerase chain reaction (PCR) amplicons of 23 TcR genes and 10 ermR genes. Total bacterial genomic DNA was extr...

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Bibliographic Details
Published in:Environmental microbiology 2007-03, Vol.9 (3), p.703-715
Main Authors: Patterson, Andrea J, Colangeli, Roberto, Spigaglia, Patrizia, Scott, Karen P
Format: Article
Language:English
Subjects:
Animal Husbandry
Animals
Anti-Bacterial Agents - pharmacology
Bacteria - classification
Bacteria - genetics
Bacteria - isolation & purification
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Cattle
DNA Transposable Elements
DNA, Bacterial - analysis
DNA, Bacterial - isolation & purification
Drug Resistance, Bacterial - genetics
Erythromycin - pharmacology
Feces - microbiology
Oligonucleotide Array Sequence Analysis - methods
RNA, Ribosomal, 16S - genetics
Sensitivity and Specificity
Sheep
Soil Microbiology
Swine
Tetracycline Resistance - genetics
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Summary:A macroarray system was developed to screen environmental samples for the presence of specific tetracycline (TcR) and erythromycin (ermR) resistance genes. The macroarray was loaded with polymerase chain reaction (PCR) amplicons of 23 TcR genes and 10 ermR genes. Total bacterial genomic DNA was extracted from soil and animal faecal samples collected from different European countries. Macroarray hybridization was performed under stringent conditions and the results were analysed by fluorescence scanning. Pig herds in Norway, reared without antibiotic use, had a significantly lower incidence of antibiotic resistant bacteria than those reared in other European countries, and organic herds contained lower numbers of resistant bacteria than intensively farmed animals. The relative proportions of the different genes were constant across the different countries. Ribosome protection type TcR genes were the most common resistance genes in animal faecal samples, with the tet(W) gene the most abundant, followed by tet(O) and tet(Q). Different resistance genes were present in soil samples, where erm(V) and erm(E) were the most prevalent followed by the efflux type TcR genes. The macroarray proved a powerful tool to screen DNA extracted from environmental samples to identify the most abundant TcR and ermR genes within those tested, avoiding the need for culturing and biased PCR amplification steps.
ISSN:1462-2912
1462-2920
DOI:10.1111/j.1462-2920.2006.01190.x