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  • Sludge as a potential important source of antibiotic resistance genes in both the bacterial and bacteriophage fractions.

Sludge as a potential important source of antibiotic resistance genes in both the bacterial and bacteriophage fractions.

Environmental science & technology (2014-05-31)
William Calero-Cáceres, Ana Melgarejo, Marta Colomer-Lluch, Claudia Stoll, Francisco Lucena, Juan Jofre, Maite Muniesa
ABSTRACT

The emergence and prevalence of antibiotic resistance genes (ARGs) in the environment is a serious global health concern. ARGs found in bacteria can become mobilized in bacteriophage particles in the environment. Sludge derived from secondary treatment in wastewater treatment plants (WWTPs) constitutes a concentrated pool of bacteria and phages that are removed during the treatment process. This study evaluates the prevalence of ARGs in the bacterial and phage fractions of anaerobic digested sludge; five ARGs (blaTEM, blaCTX-M, qnrA, qnrS, and sul1) are quantified by qPCR. Comparison between the wastewater and sludge revealed a shift in the prevalence of ARGs (blaTEM and sul1 became more prevalent in sludge), suggesting there is a change in the bacterial and phage populations from wastewater to those selected during the secondary treatment and the later anaerobic mesophilic digestion of the sludge. ARGs densities were higher in the bacterial than in the phage fraction, with high densities in both fractions; particularly for blaTEM and sul1 (5 and 8 log10 gene copies (GC)/g, respectively, in bacterial DNA; 5.5 and 4.4 log10 GC/g, respectively, in phage DNA). These results question the potential agricultural uses of treated sludge, as it could contribute to the spread of ARGs in the environment and have an impact on the bacterial communities of the receiving ecosystem.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Ampicillin, meets USP testing specifications
Ampicillin, anhydrous, European Pharmacopoeia (EP) Reference Standard
Supelco
Sulfamethoxazole, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
Nalidixic acid, ≥98%
Sigma-Aldrich
β-D-Allose, rare aldohexose sugar
Supelco
Sulfamethoxazole
Sigma-Aldrich
Ampicillin, anhydrous, 96.0-102.0% (anhydrous basis)
Supelco
Nalidixic acid, analytical standard
Sulfamethoxazole, European Pharmacopoeia (EP) Reference Standard
USP
Ampicillin, United States Pharmacopeia (USP) Reference Standard
USP
Sulfamethoxazole, United States Pharmacopeia (USP) Reference Standard
Supelco
Sulfamethoxazole, certified reference material, TraceCERT®, Manufactured by: Sigma-Aldrich Production GmbH, Switzerland
Supelco
Ampicillin, analytical standard
Supelco
Sulfamethoxazole, VETRANAL®, analytical standard