Expression of resistance genes instead of gene abundance are correlated with trace levels of antibiotics in urban surface waters.

In this study, antibiotic resistance to macrolide-lincosamide-streptogramin B (MLSB) antibiotics in total microbial community in surface water in a coastal urban city was measured using a modified fluorescence in situ hybridization (FISH) technique. This FISH technique quantified the rate of antibiotic resistance to MLSB antibiotics through targeting methylation site of A2058 of 23S rRNAs resulting from expressed erythromycin ribosome methylation (erm) genes. Correlations between the rates of MLSB resistance measured by FISH and macrolide concentrations was stronger than that between the relative abundance of erm genes and macrolide concentrations, especially in residential areas where the main detected antibiotics were macrolides. These results suggest that trace levels of antibiotics in environmental waters, which was as low as 40 ng L-1, may still play important roles in the development and spread of antibiotic resistance. Additionally, methylation as a result of erm gene expression, instead of erm gene abundance, was a better indicator of selective pressure of trace level macrolides. The rates of MLSB resistance varied significantly among land use types, suggesting that anthropogenic activities are important factors to select for erm gene expression in the environment. Microbial community analysis of representative surface water samples showed that relatively high rates of MLSB resistance were observed in Alphaproteobacteria (42%), Acidobacteria (36%), Bacteroidaceae (32%), Chloroflexi (27%), and Betaproteobacteria (20.2%).

Effects of Swine manure on macrolide, lincosamide, and streptogramin B antimicrobial resistance in soils.

Current agricultural practices involve inclusion of antimicrobials in animal feed and result in manure containing antimicrobials and antimicrobial-resistant microorganisms. This work evaluated the effects of land application of swine manure on the levels of tetracycline, macrolide, and lincosamide antimicrobials and on macrolide, lincosamide, and streptogramin B (MLS(B)) resistance in field soil samples and laboratory soil batch tests. MLS(B) and tetracycline antimicrobials were quantified after solid-phase extraction using liquid chromatography-tandem mass spectrometry. The prevalence of the ribosomal modification responsible for MLS(B) resistance in the same samples was quantified using fluorescence in situ hybridization. Macrolide antimicrobials were not detected in soil samples, while tetracyclines were detected, suggesting that the latter compounds persist in soil. No significant differences in ribosomal methylation or presumed MLS(B) resistance were observed when amended and unamended field soils were compared, although a transient (<20-day) increase was observed in most batch tests. Clostridium cluster XIVa accounted for the largest fraction of resistant bacteria identified in amended soils. Overall, this study did not detect a persistent increase in the prevalence of MLS(B) resistance due to land application of treated swine manure.

Presence of macrolide-lincosamide-streptogramin B and tetracycline antimicrobials in swine waste treatment processes and amended soil.

Little is known about the fate of antimicrobials during common agricultural waste handling procedures. To better define the potential scope of this problem, concentrations of antimicrobials throughout the waste treatment process were estimated based on known antimicrobial usage, and the resulting predictions of high antimicrobial concentrations indicated the need for further investigation. Samples from building pits, a solids settling basin, a holding pond, and soil amended with waste treatment byproducts were therefore analyzed for traditional chemical parameters and macrolide, lincosamide, and tetracycline antimicrobials. Substantial improvements in water quality were observed during the treatment process. While the macrolide tylosin was not detected, chlortetracycline, oxytetracycline, and lincomycin were found at high concentrations throughout the waste treatment process. Oxytetracycline and lincomycin were also detected in soil from a field amended with waste treatment byproducts.