A Fast and Easily Parallelizable Biosensor Method for Measuring Extractable Tetracyclines in Soils

Abstract

Quantification of extractable antibiotics in soils is important to assessing their bioavailability and mobility, and ultimately their ecotoxicological and health risks. This study aimed to establish a biosensor method for detecting extractable tetracyclines in soils (Alfisol, Mollisol, and Ultisol) using whole-cell biosensors containing a reporter plasmid (pMTGFP or pMTmCherry) carrying fluorescent protein genes tightly controlled by tetracyclines-responsive control region (<i>tet</i>RO). This whole-cell biosensor method can simultaneously measure 96 or more samples within 6 h and is easily parallelizable, whereas a typical high-performance liquid chromatography (HPLC) method may require 7 times more of analysis time and much greater cost to achieve similar analytical throughput. The biosensor method had a detection limit for each of six tetracyclines between 5.32-10.2 μg/kg soil, which is considered adequate for detecting tetracyclines in ethylenediaminetetraacetic acid (EDTA) extracts of soils. Relative standard deviation was between 19.8-51.2% for the biosensor <i>Escherichia coli</i> DH5α/pMTGFP and 2.98-25.8% for <i>E. coli</i> DH5α/pMTmCherry, respectively, suggesting that <i>E. coli</i> DH5α/pMTmCherry was superior to <i>E. coli</i> DH5α/pMTGFP for detecting extractable tetracyclines in soils. This new, fast, easily parallelizable, and cost-effective biosensor method has the potential for measuring extractable concentrations of tetracyclines for a large number of soil samples in large-scale monitoring studies.