title: Calibration and deployment of custom-designed bioreporters for protecting biological remediation consortia from toxic shock.
abstract: We have previously described the development of a panel of site-specific lux-based bioreporters from an industrial wastewater treatment system remediating coking effluents. The Pseudomonad strains carry a stable chromosomal copy of the luxCDABE operon from Photorhabdus luminescens and display proportional responses in bioluminescence decay with increasing phenol concentration up to 800 mg l-1. In this work we describe their deployment to provide a strategic sensing network for protecting bacterial communities involved in the biological breakdown of coking effluents. This evaluation demonstrated the utility of strategic placement of reporters around heavy industry treatment systems and the reliability of the reporter strains under normal operational conditions. Mono-phenol or total phenolic variation within the treatment system accounted for>65-80% of the luminescence response. The reporters exhibited stable luminescence output during normal operations with maximum standard deviations of luminescence over time of c. 5-15% depending on the treatment compartment. Furthermore, deployment of the bioreporters over a 5-month period allowed the determination of an operational range (OR) for each reporter for effluent samples from each compartment. The OR allowed a convenient measure of toxicity effects between treatment compartments and accurately reflected a specific pollution event occurring within compartments of the treatment system. This work demonstrates the utility of genetic modification to provide ecologically relevant bioreporters, extends the sensing capabilities currently obtained through marine derived biosensors and significantly enhances the potential for in situ deployment of reporting agents.
authors.affiliation: Molecular Microbial Ecology Section, CEH Oxford, Mansfield Road, Oxford, OX1 3SR, UK.
authors.forename: Siouxsie
authors.initials: S
authors.lastname: Wiles
authors.forename: Andrew K
authors.initials: AK
authors.lastname: Lilley
authors.forename: Jim C
authors.initials: JC
authors.lastname: Philp
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Bailey
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Whiteley
mesh_terms: D001673:Biodegradation, Environmental; D015374:Biosensing Techniques / Q000379:methods*; D017930:Genes, Reporter; D005818:Genetic Engineering; D008163:Luminescent Measurements; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D019800:Phenol / Q000032:analysis* / Q000633:toxicity*; D020637:Photorhabdus / Q000235:genetics; D011548:Pseudomonadaceae / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D014865:Waste Disposal, Fluid / Q000379:methods; D014874:Water Pollutants, Chemical / Q000378:metabolism; D014877:Water Pollution, Chemical; D018508:Water Purification / Q000379:methods
chemical_list: D008164:Luminescent Proteins; D014874:Water Pollutants, Chemical; D019800:Phenol