Development of a novel genetically modified bioluminescent-bacteria-based assay for detection of fluoroquinolones in animal-derived foods

Abstract

Fluoroquinolones (FQNs) are broad-spectrum antibacterial agents widely used in animal husbandry and aquaculture. The residues and antimicrobial resistance of such antibiotics are a major public health concern. To realize multianalyte detection of FQN residues, a genetically modified bacterium, Escherichia coli pK12 harboring plasmid pRecAlux3, was constructed in this study to develop a bioluminescent-bacteria-based assay for the detection of FQNs in animal-derived foods. This assay was based on the principle of induction of an SOS response by FQNs via inducing the recA-promoter-fused luciferase reporter gene existing on the plasmid pRecAlux3. E. coli pK12 was able to recognize 11 FQNs: difloxacin, enrofloxacin, ciprofloxacin, sarafloxacin, norfloxacin, danofloxacin, ofloxacin, pefloxacin, lomefloxacin, marbofloxacin, and orbifloxacin. This method could be applied to 11 edible tissues, including milk, fish muscle, and the muscles, livers, and kidneys of cattle, chickens, and pigs, with a very simple and rapid sample extraction procedure using only phosphate-buffered saline. The limits of detection of the FQNs were between 12.5 and 100 μg kg(-1), all of which were lower than the maximum residue limits. Most of the recoveries of the FQNs were in the range from 60 to 120 %, and the interassay coefficients of variation were less than 30 %. This method, confirmed by high-performance liquid chromatography, is reliable and can be used as both a screening test and a semiquantitative assay, when the identity of a single type of FQN is known.

Pharmacokinetic and Pharmacodynamic Evaluation of Marbofloxacin and PK/PD Modeling against Escherichia coli in Pigs

“…However, due to the overuse and misuse of antimicrobial agents, the resistance of the pathogenic bacterium to fluoroquinolone or other drugs was increasingly more serious ( Burgess, 1999 ). An increasing number of studies about E. coli resistance to fluoroquinolone including MBF have been reported in most countries, such as Europe, the United States, China, and so on ( Liu et al, 2013 , 2015 ; Cheng et al, 2014 ; El Garch et al, 2017 ). The cross-resistance to similar antibiotics was widespread, and the resistance mechanisms were complex.…”

Structure, Solubility and Stability of Orbifloxacin Crystal Forms: Hemihydrate versus Anhydrate

“…Fluoroquinolones are a class of antimicrobials that are widely used in human and veterinary medicine [1,2,3]. This group of antimicrobial agents has significant bactericidal activity against a wide range of veterinary pathogens and Gram-negative bacteria ( Escherichia coli , certain species of Enterobacter , Klebsiella , Pasteurella , Proteus , Salmonella , and Pseudomonas ) and some Gram-positive bacteria, such as Staphylococcus spp., Chlamydia , Mycobacteria , Mycoplasma , and Ureaplasma spp.…”

Molecularly imprinted polymers for the solid‐phase extraction of four fluoroquilones from milk and lake water samples

“…Due to the extensive application of FQs in human medicine and animal feed, the residue of the FQs is easy to cause the antimicrobial resistance. Therefore, it is important to detect FQs in animal derived food and environmental samples .…”