Antibiotics

Antibiotics belonging to the tetracycline family are extensively used in the therapy and prophylactic control of bacterial infections in human and veterinary medicine and as food additives for growth promotion in the farming industry. Intensive use of tetracyclines has led to widespread antibiotic resistance in bacterial species. The resistance mechanism genes are located in plasmids that can be efficiently transferred from one strain to another. In addition to the development of drug-resistant pathogens, the use of tetracycline has been associated with problems such as unacceptable levels of drug residues in food products for human consumption and release of drugs into the environment. Control of usage in animal farming is possible by monitoring antibiotic residues in different biological samples. Conventional methods for the detection of tetracycline residues include microbial inhibition tests, immunoassays, and chromatographic methods. Recently, whole-cell sensing systems have been proposed as a sensitive, simple, fast, and inexpensive method for measuring tetracyclines in biological fluids and food samples [111].

A bioluminescent bacterial strain containing the bacterial luciferase operon luxCDABE under the control of the tetracycline-responsive element, which is part of the regulation unit of the tetracycline resistance factor, has been developed.

It allows for the specific detection of different clinically relevant tetracyclines, with limits of detection at picomole levels and an induction time of 90 min [112]. Moreover, freeze-dried cells can detect tetracyclines as sensitively as freshly cultivated cells, thus envisioning the possibility of on-site use. The developed whole-cell sensing system has been tested on different biological samples. It has proved to be suitable to screen veterinary serum samples for tetracycline residues in real time [113] and to detect tetracyclines in raw bovine milk samples below the official limits set by the European Union and the U.S. Food and Drug Administration [114]. The biosensing strain was also applied to the detection of traces of tetracyclines in fish, after simple sample preparation. In this case, tetracycline residues were detected below official limits, and the results correlated with those obtained by conventional HPLC [115].

In another study, a GFP-based whole-cell sensor system for in situ detection of tetracyclines was developed. The biosensor was used for qualitative detection of oxytetracycline production by the bacterium Streptomyces rimosus in soil microcosms [116]. The tetracycline-induced GFP-producing biosensor cells were detected by using flow cytometry and fluorescence-activated cell-sorting (FACS) analysis. This study showed the biosensor potential for microbial ecology studies. The same sensor strain was employed in a separate study for in vivo detection and quantification of tetracyclines in rat intestine [117]. Bioavailable tetracycline concentration within the bacterial growth habitat of the intestine proved to be proportional to the intake concentration, but significantly lower. This finding, made possible by the use of the whole-cell sensing system, may help to clarify and optimize antimicrobial therapy in the intestinal environment.

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